OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
Rapidly Evolving
Genes and Genetic
Systems
EDITED BY
Rama S. Singh
McMaster University, Canada
Jianping Xu
McMaster University, Canada
and
Rob J. Kulathinal
Temple University, USA
1
Rapidly Evolving Genes and Genetic Systems. First Edition. Edited by Rama S. Singh, Jianping Xu,
and Rob J. Kulathinal. ? 2012 Oxford University Press. Published 2012 by Oxford University Press.
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
3
Great Clarendon Street, Oxford, OX2 6DP,
United Kingdom
Oxford University Press is a department of the University of Oxford.
It furthers the University?s objective of excellence in research, scholarship,
and education by publishing worldwide. Oxford is a registered trade mark of
Oxford University Press in the UK and in certain other countries
? Oxford University Press 2012
The moral rights of the authors have been asserted
First Edition published in 2012
Impression: 1
All rights reserved. No part of this publication may be reproduced, stored in
a retrieval system, or transmitted, in any form or by any means, without the
prior permission in writing of Oxford University Press, or as expressly permitted
by law, by licence or under terms agreed with the appropriate reprographics
rights organization. Enquiries concerning reproduction outside the scope of the
above should be sent to the Rights Department, Oxford University Press, at the
address above
You must not circulate this work in any other form
and you must impose this same condition on any acquirer
British Library Cataloguing in Publication Data
Data available
Library of Congress Cataloging in Publication Data
Library of Congress Control Number: 2012937854
ISBN 978?0?19?964227?4 (hbk)
978?0?19?964228?1 (pbk)
Printed and bound by
CPI Group (UK) Ltd, Croydon, CR0 4YY
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
Contents
Foreword xiv
Richard Lewontin
Preface xvi
List of Contributors xvii
1 Introduction 1
Rama S. Singh, Jianping Xu, and Rob J. Kulathinal
1.1 A gradualist history 1
1.2 Mechanisms of rapid and episodic change 2
1.2.1 Unconstrained neutral space 2
1.2.2 Horizontal gene transfer 3
1.2.3 Developmental macromutations 3
1.2.4 Evolution by gene regulation 3
1.2.5 Coevolutionary forces 4
1.2.6 Sexual selection and sexual arms races 4
1.2.7 Population demography and genetic revolutions 5
1.2.8 Adaptive radiation 5
1.3 Punctuated equilibrium within a microevolution framework 5
1.4 Tempo, mode, and the genomic landscape 6
1.5 ?Rapidly evolving genes and genetic systems?: a brief overview 7
1.6 Future prospects 8
Part I From Theory to Experiment
2 Theoretical perspectives on rapid evolutionary change 13
Sarah P. Otto
2.1 Introduction 13
2.2 When is strong selection strong? 13
2.3 Does strong selection differ in kind from weak selection? 16
2.4 Concluding thoughts 20
3 Recombination reshuffles the genotypic deck, thus accelerating the rate
of evolution 23
Mihai Albu, Amir R. Kermany, and Donal A. Hickey
3.1 Introduction 23
3.2 Simulating selection on multilocus genotypes 24
3.3 Discussion 27
3.4 Conclusions 29
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
vi CONTENTS
4 Heterogeneity in neutral divergence across genomic regions induced by
sex-specific hybrid incompatibility 31
Seiji Kumagai and Marcy K. Uyenoyama
4.1 Introduction 31
4.1.1 Detecting incompatibility factors 31
4.1.2 Within-species polymorphisms for incompatibility factors with
sex-limited transmission 31
4.2 Genealogical migration rate 32
4.2.1 Definition 32
4.2.2 Non-sex-specific incompatibility 33
4.2.3 Sex-specific incompatibility 33
4.3 Applications 33
4.3.1 Mitochondrial introgression 33
4.3.2 Interpreting region-specific FST 35
4.4 Conclusions 37
5 Rapid evolution in experimental populations of major life forms 40
Jianping Xu
5.1 Introduction 40
5.2 Features of experimental evolution 41
5.3 Types of experimental evolution 42
5.3.1 Directional selection 42
5.3.2 Adaptation 42
5.3.3 Mutation accumulation 42
5.4 Rapid change and divergence among mutation accumulation
population lines 43
5.4.1 Microbial growth rate 43
5.4.2 Other microbial traits 45
5.4.3 Plants and animals 45
5.5 Adaptation and directional selection experiments 47
5.5.1 Adaptation of E. coli populations 47
5.5.2 Adaptation of viral populations 47
5.5.3 Adaptation and directional selection in fruit flies 48
5.5.4 Adaptation in yeast 48
5.5.5 Directional selection in mammals 48
5.5.6 Correlated changes between traits 49
5.5.7 Acquisition of novel phenotypes 49
5.6 Genomic analysis of experimental evolution populations 50
5.7 Conclusions and perspectives 50
Part II Rapidly Evolving Genetic Elements
6 Rapid evolution of low complexity sequences and single amino
acid repeats across eukaryotes 55
Wilfried Haerty and G. Brian Golding
6.1 Introduction 55
6.2 Rapid evolution of low complexity sequences 55
6.2.1 Mutational processes 56
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
CONTENTS vii
6.3 Rapid divergence of LCRs and their impact on surrounding sequences 57
6.3.1 LCRs as indicators of regions of lowered purifying
selective pressures 57
6.3.2 Mutagenic effect of LCRs 58
6.4 Low complexity sequences under selection 59
6.4.1 Deleterious effects of LCR size variation 59
6.4.2 DNA composition 59
6.4.3 LCR distribution 60
6.4.4 Phenotypic effects of LCR size variation 60
6.4.5 Selection for low information content 61
6.5 Perspectives 61
7 Fast rates of evolution in bacteria due to horizontal gene transfer 64
Weilong Hao
7.1 Introduction 64
7.2 Quantifying horizontal gene transfer 65
7.3 Understanding the variation of gene gain and loss 66
7.4 Horizontal gene transfer in duplicated genes 67
7.5 Pseudogenization of horizontally transferred genes 67
7.6 Mobile sequences and gene movement 68
7.7 Gene exchange goes fine-scale 69
7.8 Conclusions 69
8 Rapid evolution of animal mitochondrial DNA 73
Xuhua Xia
8.1 Introduction 73
8.2 Mitochondrial replication, strand bias, and evolutionary rates 74
8.3 The change in genetic code and evolutionary rate 77
8.4 The change in tRNA genes and evolutionary rate 79
8.5 Conclusions 81
9 Rapid evolution of centromeres and centromeric/kinetochore proteins 83
Kevin C. Roach, Benjamin D. Ross, and Harmit S. Malik
9.1 Centromeres in ?the fast lane? 83
9.2 Rapidly evolving centromeric histones 83
9.3 Bewildering centromeric DNA complexity and evolution 85
9.4 The ?centromere paradox?: conflict, not coevolution 87
9.5 Support for the centromere drive model 89
9.6 Taxonomic differences in susceptibility to centromere drive 89
9.7 Rapid evolution of other centromeric proteins 90
9.8 Centromere drive and postzygotic isolation between species 91
9.9 Future directions 91
10 Rapid evolution via chimeric genes 94
Rebekah L. Rogers and Daniel L. Hartl
10.1 Introduction 94
10.2 Mechanisms of formation 94
10.3 Selection 96
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
viii CONTENTS
10.4 Genomic stability 96
10.5 Function 97
10.6 Non-coding DNA 98
10.7 Future directions 99
11 Evolutionary interactions between sex chromosomes and autosomes 101
Manyuan Long, Maria D. Vibranovski, and Yong E. Zhang
11.1 Introduction 101
11.2 Gene traffic between sex chromosome and autosomes 102
11.2.1 Gene traffic in Drosophila 102
11.2.2 Gene traffic in mammals 103
11.2.3 The cause and consequence of gene traffic 104
11.3 The generality of gene traffic out of the X in the genus Drosophila 105
11.3.1 Gene traffic in Drosophilidae and RNA-based and DNA-based
duplication 105
11.3.2 Independent tests of gene traffic 105
11.4 Mechanisms underlying gene traffic out of the X: the detection of meiotic
sex chromosome inactivation 107
11.4.1 Evolutionary genetic models 107
11.4.2 Molecular mechanistic models 107
11.5 The X-recruitment of young male-biased genes and gene traffic out of the
X chromosome 108
11.5.1 Age-dependence in Drosophila 109
11.5.2 Age-dependence in mammals 110
11.5.3 The slow enrichment of X-linked female genes 110
11.6 Concluding remarks 111
12 Evolutionary signatures in non-coding DNA 115
Dara G. Torgerson and Ryan D. Hernandez
12.1 Introduction 115
12.2 Challenges to studying the evolution of non-coding DNA 116
12.2.1 Identifying functional non-coding DNA 116
12.2.2 Estimating the neutral evolutionary rate 117
12.2.3 Limitations of identifying rapid evolution in non-coding DNA 117
12.3 Patterns of evolution in non-coding DNA 117
12.3.1 Selection in conserved non-coding sequences? 118
12.3.2 Detecting selection in promoters and TFBSs 120
12.3.3 Emerging trends in microRNA binding sites 121
12.3.4 Coding versus non-coding 121
12.4 Future prospects 122
Part III Sex- and Reproduction-Related Genetic Systems
13 Evolution of sperm?egg interaction 127
Melody R. Palmer and Willie J. Swanson
13.1 Introduction 127
13.2 Evolution at each step of sperm?egg interaction 127
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
CONTENTS ix
13.3 Causes of rapid evolution 130
13.4 Methods to identify interacting proteins 132
13.5 Conclusions 132
14 Rates of sea urchin bindin evolution 136
H. A. Lessios and Kirk S. Zigler
14.1 Introduction 136
14.2 Function and structure of bindin 136
14.3 Rate of bindin evolution 137
14.4 Possible reasons for different evolutionary rates in bindin 139
14.5 Conclusions and future prospects 141
15 Evolution of Drosophila seminal proteins and their networks 144
Alex Wong and Mariana F. Wolfner
15.1 Introduction 144
15.2 Drosophila seminal fluid as a model system for rapidly
evolving proteins 144
15.3 Extensive variation in rates of SFP evolution 147
15.4 Selection on a network? 149
15.5 Conclusions 150
16 Evolutionary genomics of the sperm proteome 153
Timothy L. Karr and Steve Dorus
16.1 Introduction 153
16.2 Characterization of the Drosophila sperm proteome 154
16.3 Molecular evolution of the Drosophila sperm proteome 154
16.4 Evolution of novel Drosophila sperm components 155
16.4.1 Novel genes in the sperm proteome 156
16.4.2 Expansion and diversification of S-LAP gene family 157
16.5 The mouse sperm proteome: intensified selection on sperm membrane
and acrosome genes 157
16.6 Rapid evolution of immunity-related genes in mammalian sperm 160
16.7 Sexual selection and compartmentalized adaptation in reproductive
genetic systems 161
16.8 Future perspectives 162
17 Fast evolution of reproductive genes: when is selection sexual? 165
Alberto Civetta
17.1 Introduction 165
17.2 What has been the role of selection during the evolution of male
reproductive genes? 167
17.3 When is selection sexual? The phylogenetic approach 168
17.4 Testing sexual selection in the era of genomes 168
17.5 The need for association studies and functional assays 171
17.6 Conclusions 172
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
x CONTENTS
18 Rapid morphological, behavioral, and ecological evolution in Drosophila:
comparisons between the endemic Hawaiian Drosophila and the
cactophilic repleta species group 176
Patrick M. O?Grady and Therese Ann Markow
18.1 Introduction 176
18.1.1 Ecological adaptations 177
18.1.2 Morphological adaptations 177
18.1.3 Behavioral adaptations 178
18.2 Hawaiian Drosophila radiation 179
18.2.1 Phylogenetic relationships 179
18.2.2 Sexual adaptations to morphology and behavior 179
18.2.3 Ecological adaptations to morphology and behavior 179
18.3 Cactophilic Drosophila radiation in the New World 180
18.3.1 Phylogenetic relationships 180
18.3.2 Rapid evolution of ecological adaptations 180
18.3.3 Rapid evolution of behavioral traits 182
18.4 Conclusions: adaptive radiation versus adaptive infiltration 183
19 Ancient yet fast: rapid evolution of mating genes and mating systems
in fungi 187
Timothy Y. James
19.1 Introduction 187
19.2 Incompatibility systems in fungi 189
19.3 Fungal reproductive proteins show evidence for positive and
balancing selection 190
19.4 Evidence for rapid evolution of fungal incompatibility genes
and systems 193
19.4.1 Sequence evolution 194
19.4.2 Mating systems and loci 194
19.5 Evidence for ancient alleles and mating systems 196
19.6 Conclusions 198
Part IV Pathogens and their Hosts
20 Rapid evolution of innate immune response genes 203
Brian P. Lazzaro and Andrew G. Clark
20.1 The evolution of immunity 203
20.2 Orthology and gene family evolution in antimicrobial immunity 204
20.3 Molecular evolution of the antimicrobial immune system 205
20.4 The evolution of defense against viruses and transposable elements 206
20.5 Concluding remarks 208
21 Rapid evolution of the plague pathogen 211
Ruifu Yang, Yujun Cui, and Dongsheng Zhou
21.1 Introduction 211
21.2 Plasmid acquisition in Y. pestis 212
21.3 The impact of phages on genome structure 213
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
CONTENTS xi
21.4 Prophages in the Y. pestis genome 213
21.5 CRISPRs diversity and the battle between phage and Y. pestis 214
21.6 Gene acquisition, loss, and inactivation 216
21.7 Rearrangements and copy number variants 217
21.8 Neutral versus adaptive evolution 219
21.9 Conclusions 220
22 Evolution of human erythrocyte-specific genes involved in malaria
susceptibility 223
Wen-Ya Ko, Felicia Gomez, and Sarah A. Tishkoff
22.1 Introduction 223
22.2 Adaptive evolution in erythrocyte-specific genes 224
22.2.1 Genetic variants causing erythrocytic structural, regulatory, or
enzymatic deficiency: candidates for heterozygote advantage 224
22.2.2 Positive selection on erythrocyte-surface receptors 226
22.3 Evolutionary response of the human genome to malaria infection 227
22.3.1 Maintenance of deleterious mutations due to selective
pressure of malaria 227
22.3.2 Effects of population substructure on genetic variation in
malaria-endemic human populations 230
22.3.3 Effects of gene conversion between homologous sequences on
genetic variation at loci associated with malarial susceptibility 232
22.4 Future perspectives 232
Part V From Gene Expression to Development to Speciation
23 The rapid evolution of gene expression 237
Carlo G. Artieri
23.1 Introduction 237
23.2 One genome harbors many transcriptomes 238
23.3 Transcriptome divergence is complex 239
23.4 Factors affecting the rate of evolution of gene expression 240
23.4.1 Spatial heterogeneity 240
23.4.2 Temporal heterogeneity 241
23.5 Beyond comparisons of expression levels 242
23.6 Open questions and future directions 243
24 Rate variation in the evolution of development: a phylogenetic
perspective 246
Artyom Kopp
24.1 Introduction 246
24.2 Examples of rate variation in the evolution of development 247
24.2.1 Same clade, different pathways: evolution of vulval development
in rhabditid nematodes 247
24.2.2 Same pathway, different clades: evolution of sex combs and
pigmentation in Drosophila 248
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
xii CONTENTS
24.2.3 Same clade, same pathway, different genes: evolution of
embryonic development and sex determination in insects 251
24.3 Technical and conceptual challenges to quantifying the evolution
of development 252
24.4 Future directions: the promise of phylogenetic approaches to the
evolution of development 253
25 Natural hybridization as a catalyst of rapid evolutionary change 256
Michael L. Arnold, Jennafer A.P. Hamlin, Amanda N. Brothers, and
Evangeline S. Ballerini
25.1 Introduction 256
25.2 Adaptive trait introgression: when strange is really good 256
25.2.1 Adaptive trait transfer in Canis: wolves in dogs? clothing 257
25.2.2 Adaptive trait origin in Saccharomyces cerevisiae: hybrids
make the best wine 258
25.3 Hybrid speciation: when opposites attract 259
25.3.1 Homoploid hybrid speciation: hybrid butterflies (quickly)
change their spots 259
25.3.2 Allopolyploid speciation: Tragopogon hybrid polyploids form
again, and again, and again . . . in less than 100 years . . . 260
25.4 Natural hybridization and adaptive radiations: hybrid speciation
on steroids 261
25.4.1 Hybridization and adaptive radiations of Lake Malawi cichlids:
from hybrid swarm to 800 species, in one lake?! 261
25.4.2 Hybridization and adaptive radiations in Alpine lake whitefish:
Swiss fish diversify after the last big thaw 262
25.4.3 Hybridization and adaptive radiations in Hawaiian silverswords:
allopolyploids in an island paradise 263
25.5 Conclusions and future prospects 264
26 Rapid evolution of pollinator-mediated plant reproductive isolation 266
Annika M. Moe, Wendy L. Clement, and George D. Weiblen
26.1 Plant?insect diversification 266
26.2 Pollination and reproductive isolation 266
26.3 Ficus versus Castilleae 267
26.4 A pollinator-mediated model for fig speciation 269
26.5 Future directions: plant?pollinator interactions and rapid evolution 271
27 Sexual system genomics and speciation 274
Rob J. Kulathinal and Rama S. Singh
27.1 In the beginning: Darwin and Wallace on sexual selection and speciation 274
27.2 The Modern Synthesis and the development of speciation theory 275
27.3 A new paradigm: the genomics of sexual systems and the origin
of species 276
27.3.1 Functional genomics: organization into sexual and
non-sexual systems 277
27.3.2 Higher variation among reproductive systems 277
OUP CORRECTED PROOF ? FINAL, 29/5/2012, SPi
CONTENTS xiii
27.3.3 Strength of sexual selection 278
27.3.4 Sexual systems interaction, coevolution, and rapid change 279
27.3.5 Rapid breakdown of sexual systems in species hybrids 280
27.4 Towards a post-genomics synthesis of speciation 280
27.5 Future prospects: sex as a major force in evolution 281
Index 285
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
CHAPTER 14
Rates of sea urchin bindin evolution
H. A. Lessios and Kirk S. Zigler
14.1 Introduction
Reproduction at the level of gametic interactions
involves activation and attraction of the sperm by
egg compounds, induction of the acrosome reaction
by the egg jelly, adhesion of the sperm to the egg,
and fusion of the two membranes in order to permit
the transmission of genetic material. All of these
interactions are mediated by molecules. Some of
these molecules, such as sea urchin speract, carry
out their functions indiscriminately, even if sperm
and egg belong to distantly related taxa (Vieira
and Miller 2006). Others function in a species-
specific or even genotype-specific manner. Selectiv-
ity between sperm gamete recognition molecules
and their egg receptors is particularly important
in organisms with external fertilization, because
in the absence of copulation, there are few other
opportunities for exercising mate choice. Conse-
quently, such molecules are exposed to the action
of selection more directly than molecules with the
same function in organisms with internal fertiliza-
tion. The DNA that codes for gamete recognition
molecules often, but not always, evolves rapidly,
displaying ratios of amino acid replacement to
synonymous substitutions larger than unity, a sig-
nature of positive (diversifying) selection (Swan-
son and Vacquier 2002a, b; Vacquier and Swanson
2011). As a rule, such positive selection is targeted
at certain regions of each molecule, presumably
involved in gamete selectivity, whereas the rest
of the sequence may evolve conservatively under
purifying selection, because it performs basic func-
tions essential for fertilization.
The first gamete recognition protein to be charac-
terized was sea urchin bindin (Vacquier and Moy
1977). Bindin DNA was subsequently amplified
and sequenced in Strongylocentrotus purpuratus by
Gao et al. (1986), and then studied with regards
to its intra- and interspecific polymorphism with
special attention given to detecting positive selec-
tion in its exons. These topics have been exten-
sively reviewed (Vacquier et al. 1995; Swanson and
Vacquier 2002a, b; Lessios 2007, 2011; Zigler 2008;
Palumbi 2009; Vacquier and Swanson 2011). In this
chapter, we explore what bindin sequences from
various sea urchin species reveal about the rate
of evolution of this molecule. Does bindin really
evolve in the fast lane?
14.2 Function and structure of bindin
Sea urchin bindin is a protein that coats the acro-
some process of sperm after the acrosomal reaction
occurs. It interacts with the egg bindin receptor,
EBR1, a glycoprotein (Kamei and Glabe 2003), to
attach the sperm to the egg?s vitelline layer and
to fuse membranes of the gametes. The full-length
precursor of bindin is cleaved after translation to
form the mature molecule. Among the sea urchin
species that have been studied to date, the length
of mature bindin ranges from 193?418 amino acids
(Zigler and Lessios 2003a). The single sea star in
which bindin has been characterized was found
to contain 793 amino acids (Patino et al. 2009). In
both sea urchins and sea stars, there is a single
intron separating two exons. Bindins of 11 species
of sea urchins from six orders contain a conserved
region in the second exon that codes for approxi-
mately 55 amino acids. Eighteen amino acids in this
conserved region, thought to be involved in mem-
brane fusion (Rocha et al. 2008), have not changed
since the extant orders of Echinoidea split from
each other, 250 million years ago (mya). Only one
amino acid in this region has changed between sea
stars and sea urchins in the 500 million years (my)
Rapidly Evolving Genes and Genetic Systems. First Edition. Edited by Rama S. Singh, Jianping Xu, and Rob J. Kulathinal.
? 2012 Oxford University Press. Published 2012 by Oxford University Press.
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
RATES OF SEA URCHIN BINDIN EVOLUTION 137
that the two echinoderm classes have been evolv-
ing independently (Patino et al. 2009; Vacquier and
Swanson 2011). The reputation of bindin as a fast-
evolving protein is owed to two regions flanking
the conserved core, which in some genera have
accumulated many point mutations and insertions?
deletions. These are the regions that most likely
confer fertilization species-specificity (Lopez et al.
1993). The protein moiety of EBR1, which contains
3713?4595 amino acids, has only been sequenced in
two species of Strongylocentrotus (Kamei and Glabe
2003).
14.3 Rate of bindin evolution
Bindin has been sequenced in 11 genera of sea
urchins, but intrageneric variation, which permits
insights in the evolution of the molecule, has
been studied in only seven: Echinometra (Metz
and Palumbi 1996; McCartney and Lessios 2004),
Strongylocentrotus (Biermann 1998), Arbacia (Metz
et al. 1998a), Tripneustes (Zigler and Lessios 2003b),
Heliocidaris (Zigler et al. 2003), Lytechinus (Zigler
and Lessios 2004), and Paracentrotus (Calderon et al.
2009, 2010). Selection on bindin in all of these genera
has been studied as the ratio of amino acid replace-
ment to silent substitutions (? = dN/dS). By this cri-
terion, there is evidence of positive selection (? >1)
in Echinometra, Strongylocentrotus, Heliocidaris, and
Paracentrotus, but not in Arbacia, Tripneustes, and
Lytechinus. In addition to being an indication of
selection at the nucleotide level, the ? ratio would
also be a good measure of relative rates of adap-
tive evolution if silent sites evolved at the same
rate in all genera. This, however, is not the case
in bindin. Bindins with higher rates of nonsynony-
mous substitution also have higher rates of syn-
onymous substitution (Zigler and Lessios 2003b).
This correlation has also been observed in other
molecules such as alcohol dehydrogenase, ATP syn-
thetase, cyclophilin 1, or enolase (e.g. Dunn et al.
2001), and there are a number of hypotheses as to
its cause. While it is typically thought to arise from
some form of codon bias, codon usage in sea urchin
bindin is very equitable (Zigler and Lessios 2003a).
Thus, due to different codon biases, comparing ?
ratios between bindins of different genera may lead
to erroneous conclusions regarding evolutionary
rates. To compare the absolute rate of evolution
between genera we need to determine the number
of nonsynonymous substitutions per nonsynony-
mous site that accumulate per unit time. Such a cal-
culation requires evidence of dates of divergence. In
this chapter, we will use the interspecific divergence
of cytochrome oxidase I (COI) as a proxy for the
time since speciation. Calibrated by the rise of the
Isthmus of Panama, approximately 3 mya, COI of
sea urchins diverges at an average rate of 3.6 % per
my (Lessios 2008).
Gauged by divergence in COI, average rates
of adaptive divergence of bindin within a genus
vary between 2.80 ? 10?3 nonsynonymous substi-
tutions per nonsynonymous site per my (dNmy?1)
in Arbacia and 22.4 ? 10?3 dNmy?1 in Strongylocen-
trotus (Table 14.1). As one might expect, genera
in which bindin evolves under positive selection,
show amino acid divergence rates almost four times
higher than genera in which bindin appears to be
under purifying selection: the average substitution
rate in Strongylocentrotus, Echinometra, and Helioci-
daris is 20.4 ? 10?3 dNmy?1 whereas in Arbacia, Trip-
neustes, Lytechinus, Pseudoboletia, and Diadema, it is
5.96 ? 10?3 dNmy?1. The question we would like
to answer is how these rates of adaptive evolution
compare with those of other proteins, both of those
that have been deemed to evolve rapidly in other
taxa, and those that carry out other functions in sea
urchins.
Fig. 14. 1 presents a comparison of the rates of
adaptive evolution of bindin to seven other classes
of reproductive proteins from five groups of organ-
isms. These are all proteins that are generally con-
sidered as fast-evolving. Because COI in different
taxa evolves at different rates, it is necessary to
apply taxon-specific calibrations to calculate diver-
gence rates. To estimate absolute rates of protein
evolution, we have assumed that COI evolves at
an average rate of 3.6% per my in sea urchins
(Lessios 2008), 2.7% per MY in gastropods (Lessios
2008), 2.3% per my in insects (Papadopoulou et al.
2010), and 1.6% per my in hominids (Kumar et al.
2005). Estimated in this manner, the evolutionary
rates of bindins in different genera of sea urchins,
even those found to be under selection, are slower
than that of reproductive proteins of gastropods
or insects. They are more comparable to those of
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
Ta
bl
e
14
.1
Pa
irw
ise
di
ve
rg
en
ce
in
bi
nd
in
an
d
in
cy
to
ch
ro
m
e
ox
id
as
e
I(
CO
I)
of
se
le
ct
ed
sp
ec
ie
s
of
se
a
ur
ch
in
ge
ne
ra
in
w
hi
ch
bi
nd
in
va
ria
tio
n
ha
s
be
en
st
ud
ie
d.
K2
P:
Ki
m
ur
a
tw
o-
pa
ra
m
et
er
di
st
an
ce
;d
N
:
am
in
o
ac
id
su
bs
tit
ut
io
ns
pe
rn
on
-s
yn
on
ym
ou
s
sit
e;
d S
:s
yn
on
ym
ou
s
su
bs
tit
ut
io
ns
pe
rs
yn
on
ym
ou
s
sit
e;
M
Y:
m
ill
io
n
ye
ar
s.
Es
tim
at
ed
ra
te
s
of
di
ve
rg
en
ce
of
bi
nd
in
ar
e
ba
se
d
on
th
e
as
su
m
pt
io
n
th
at
CO
Ii
n
se
a
ur
ch
in
sd
ive
rg
es
at
a
ra
te
of
0.
03
6
pe
rs
ite
pe
rm
y.
Ge
nu
s
Sp
ec
ie
s
Sp
ec
ie
s
Bi
nd
in
CO
I
Bi
nd
in
d N
/
Bi
nd
in
d S
/
Bi
nd
in
d N
/
M
Y
Re
fe
re
nc
e
d N
d S
K2
P
CO
IK
2P
CO
IK
2P
Ar
ba
cia
lix
ul
a
pu
nc
tu
la
ta
0.
00
7
0.
06
9
0.
09
0
0.
07
2
0.
76
4
0.
00
26
M
et
z
et
al
.1
99
8a
Ar
ba
cia
lix
ul
a
st
el
la
ta
=i
nc
isa
0.
00
7
0.
09
6
0.
13
4
0.
05
3
0.
71
6
0.
00
19
Ar
ba
cia
lix
ul
a
du
fre
sn
ei
0.
01
6
0.
07
1
0.
12
4
0.
12
9
0.
57
0
0.
00
46
Ar
ba
cia
pu
nc
tu
la
ta
st
el
la
ta
=i
nc
isa
0.
00
3
0.
08
8
0.
13
9
0.
02
2
0.
63
5
0.
00
08
Ar
ba
cia
pu
nc
tu
la
ta
du
fre
sn
ei
0.
01
1
0.
05
9
0.
12
4
0.
08
5
0.
47
7
0.
00
31
Ar
ba
cia
st
el
la
ta
=i
nc
isa
du
fre
sn
ei
0.
01
3
0.
07
1
0.
11
9
0.
10
5
0.
59
7
0.
00
38
He
lio
cid
ar
is
er
yt
hr
og
ra
m
m
a
tu
be
rc
ul
at
a
0.
06
9
0.
14
9
0.
14
7
0.
46
9
1.
01
4
0.
01
69
Zi
gl
er
et
al
.2
00
3
Tr
ip
ne
us
te
s
ve
nt
ric
os
us
gr
at
illa
+d
ep
re
ss
us
0.
01
6
0.
02
6
0.
08
7
0.
18
7
0.
29
3
0.
00
67
Zi
gl
er
an
d
Le
ss
io
s2
00
3
Ec
hi
no
m
et
ra
ob
lo
ng
a
m
at
ha
ei
0.
02
1
0.
05
4
0.
02
3
0.
90
5
2.
32
8
0.
03
26
M
et
z
an
d
Pa
lu
m
bi
19
96
Ec
hi
no
m
et
ra
ob
lo
ng
a
ty
pe
A
0.
02
4
0.
07
6
0.
03
2
0.
75
7
2.
37
1
0.
02
73
Ec
hi
no
m
et
ra
m
at
ha
ei
ty
pe
A
0.
02
8
0.
05
1
0.
02
4
1.
16
9
2.
10
7
0.
04
21
Ec
hi
no
m
et
ra
lu
cu
nt
er
vir
id
is
0.
02
2
0.
04
7
0.
05
0
0.
44
0
0.
94
0
0.
01
58
M
cC
ar
tn
ey
an
d
Le
ss
io
s2
00
4
Ec
hi
no
m
et
ra
lu
cu
nt
er
va
nb
ru
nt
i
0.
02
6
0.
04
6
0.
10
2
0.
25
5
0.
45
1
0.
00
92
Ec
hi
no
m
et
ra
vir
id
is
va
nb
ru
nt
i
0.
01
4
0.
08
3
0.
12
6
0.
11
1
0.
65
9
0.
00
40
Ly
te
ch
in
us
pi
ct
us
va
rie
ga
tu
s
0.
01
3
0.
10
5
0.
13
5
0.
09
6
0.
77
8
0.
00
35
Zi
gl
er
an
d
Le
ss
io
s2
00
4
Ly
te
ch
in
us
va
rie
ga
tu
s
w
illi
am
si
0.
00
6
0.
02
2
0.
01
7
0.
35
3
1.
29
4
0.
01
27
Ly
te
ch
in
us
se
m
itu
be
rc
ul
at
us
pi
ct
us
0.
02
5
0.
07
3
0.
11
4
0.
21
9
0.
64
0
0.
00
79
Ly
te
ch
in
us
eu
er
ce
s
Sp
ha
er
ec
hi
nu
sg
ra
nu
la
ris
0.
01
9
0.
10
0
0.
08
9
0.
21
3
1.
12
4
0.
00
77
Ps
eu
do
bo
le
tia
in
di
an
a
m
ac
ul
at
a
0.
00
6
0.
02
4
0.
07
3
0.
08
2
0.
32
9
0.
00
30
Zi
gl
er
et
al
.(
in
pr
es
s)
St
ro
ng
ylo
ce
nt
ro
tu
s
pu
rp
ur
at
us
pa
llid
us
0.
02
1
0.
06
2
0.
07
2
0.
28
7
0.
86
3
0.
01
03
Bi
er
m
an
n
19
98
St
ro
ng
ylo
ce
nt
ro
tu
sp
ur
pu
ra
tu
s
pa
llid
us
dr
oe
ba
ch
ie
ns
is
0.
03
1
0.
08
6
0.
07
5
0.
41
8
1.
14
8
0.
01
50
St
ro
ng
ylo
ce
nt
ro
tu
s
pu
rp
ur
at
us
H.
pu
lch
er
rim
us
0.
07
3
0.
15
8
0.
10
4
0.
70
4
1.
51
4
0.
02
53
St
ro
ng
ylo
ce
nt
ro
tu
s
pa
llid
us
dr
oe
ba
ch
ie
ns
is
0.
02
5
0.
03
6
0.
03
5
0.
71
5
1.
01
1
0.
02
57
St
ro
ng
ylo
ce
nt
ro
tu
s
pa
llid
us
H.
pu
lch
er
rim
us
0.
06
6
0.
11
9
0.
07
0
0.
94
1
1.
69
6
0.
03
39
St
ro
ng
ylo
ce
nt
ro
tu
s
dr
oe
ba
ch
ie
ns
is
H.
pu
lch
er
rim
us
0.
06
3
0.
13
9
0.
09
4
0.
67
2
1.
48
1
0.
02
42
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
RATES OF SEA URCHIN BINDIN EVOLUTION 139
B
0
10
20
30
40
HL H18 TL
d
N
 /
 m
y 
?
 1
0?
2
TMAP
Selected rapidly evolving reproductive proteins
Acps P ZP/OGP
Figure 14.1 Bindin evolution relative to known fast-evolving
reproductive proteins from other taxa. Non-synonymous substitutions per
non-synonymous site (dN) per million years, between congeneric species
(except in hominids, in which they are within the same family) in sea urchin
bindin (B) (data from references in Table 14.1), abalone lysin (HL) and 18
kD protein (H18) (data from Metz et al. 1998b), Tegula lysin (TL), and the
mature region of TMAP protein (TMAP) (data from Hellberg and Vacquier
1999; Hellberg et al. 2000), Drosophila Acp26Aa and Acp36DE (Acps)
(data from Tsaur and Wu 1997), hominid protamine 1 and 2 (P), ZP2, ZP3
and oviductal glycoprotein (ZP/OGP) (data from Wyckoff et al. 2000).
protamines, zona pellucida proteins, and oviductal
glycoprotein in hominids. Adjustments to the
assumed rate of COI evolution, or even an assump-
tion of a universal COI clock, would not change
this conclusion. Thus, by the standard of other fast-
evolving reproductive proteins from other inverte-
brates, bindin evolves only at moderate rates.
How do rates of bindin evolution compare to
rates of evolution among other sea urchin pro-
teins? To answer this question, we compared all
protein coding DNA sequences of Lytechinus var-
iegatus in GenBank to their closest matches in
the Strongylocentrotus purpuratus complete genome.
With the exception of S. purpuratus, more genes
have been sequenced from Lytechinus variegatus
than any other species of sea urchin. Lytechinus
and Strongylocentrotus diverged approximately 60
mya. Sequences were available for 90 L. variegatus
genes. The protein sequence of each gene was com-
pared between the two species via protein-protein
BLAST to GenBank?s ?non-redundant (nr) protein
sequences? database. The closest match to a S. pur-
puratus protein was noted, and the two protein
sequences were aligned using Clustal in MEGA
(v. 4.0). We then used MEGA to calculate the p-
distance between the aligned protein sequences.
We identified matches for 85 of the 90 Lytechinus
genes. The five genes that did not have a match
may be: (1) missing from the annotated Strongy-
locentrotus genome; (2) lost in the Strongylocentro-
tus lineage; or (3) mis-annotated in their original
Lytechinus entry. The set of genes that we compared
contained proteins with various functions, includ-
ing many involved in reproduction, and also in
development, cytoskeleton formation, cell attach-
ment, and stress responses. After ranking the diver-
gences of the 85 proteins, that of bindin was the
sixth largest, with a p-distance of 0.326 for the full-
length molecule and 0.314 for the mature portion.
Of the five proteins with divergence values higher
than bindin, vitellogenin and SFE-1 also carry out
functions related to reproduction, whereas the other
three were involved in development. Considering
the inevitable bias of proteins available for compar-
ison, the conclusion from this comparison is that
bindin evolves at moderately fast rates in relation
to other sea urchin proteins.
14.4 Possible reasons for different
evolutionary rates in bindin
Why does bindin in four sea urchin genera evolve
more rapidly under strong positive selection, than
in three other genera in which it is subject to puri-
fying selection? In the absence of data regarding
variation in its egg receptor, the answer can only be
speculative. Possible reasons for this lack of pattern
have been thoroughly reviewed (Lessios 2007, 2011;
Zigler 2008; Palumbi 2009). Here we present a sum-
mary of the hypotheses that have been proposed
so far.
One possibility is that positive selection of
bindin arises from the need for species recogni-
tion when two closely related species are in danger
of hybridizing with each other. We will call this
the ?reinforcement hypothesis.? This name does not
imply that speciation by reinforcement has actually
taken place, but rather that bindin alleles resem-
bling those of a sympatric species?and thus allow-
ing gamete wastage in inferior hybrids?have been
selected against. A broad-brush picture of compar-
isons between genera is consistent with this hypoth-
esis. When bindin rates of divergence of species
that are entirely allopatric with respect to congeners
are compared to those of species that may have a
higher probability of hybridization, those of the for-
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
140 RAPIDLY EVOLVING GENES AND GENETIC SYSTEMS
mer are clustered around lower values than those of
the latter (Fig. 14.2). Genera with many sympatric
species, such as Strongylocentrotus, and Echinometra
tend to have the highest rates of interspecific bindin
divergence. Not all the data, however, are consis-
tent with the reinforcement hypothesis. Contrary to
what is expected from selection for species recog-
nition, bindin is polymorphic and shows the signa-
ture of positive selection not just between species
but also between alleles of the same species (Metz
and Palumbi 1996; Lessios 2007, 2011). A pattern
of character displacement is present in one species
of Pacific Echinometra (Geyer and Palumbi 2003) in
partial geographic overlap with its sister species
but not in an Atlantic species of the same genus
that also needs to contend with the challenge of a
sister species existing over part of its range (Geyer
and Lessios 2009). Given the present evidence, the
hypothesis that reinforcement in sympatry acceler-
ates bindin divergence is as likely as the hypothe-
sis that divergence in bindin, due to other causes,
allows for sympatric coexistence.
Another possibility for the differences in rates
of bindin evolution could be that they are cor-
Allopatric
Pseudoboletia
Arbacia
Strongylocentrotus S
Echinometra S
Tripneustes
Heliocidaris S
Lytechinus
0.0
0.2
0.4
B
in
d
in
 d
N
/
K
2P
C
O
I
0.6
0.8
1.0
1.2
1.4
Sympatric
Figure 14.2 Comparison of interspecific rates of bindin divergence
between genera. Amino acid replacement substitutions (dN) per
replacement site in bindin divided by Kimura-two-parameter distance in
cytochrome oxidase I (COI K2P) in allopatric and sympatric species of eight
genera of sea urchins. A species is considered as ?allopatric? if its range
does not overlap with that of another member of the same genus. Genera
in which bindin has been shown to be under selection are marked in the
legend with S.
related to the relative age of species in different
sea urchin genera. If, as Civetta and Singh (1998)
have suggested, episodes of divergence in repro-
ductive molecules are concentrated at the time
of speciation, and if selection on these molecules
is subsequently relaxed, younger species would
show higher rates of bindin differentiation than
older ones. This hypothesis is not supported by
the data. Sea urchins tend to conform to ?Jordan?s
rule? (Jordan 1905). Young sister species tend to
be distributed on either side of a geographic bar-
rier, and only older species become sympatric with
the passage of time (Lessios 2010). Thus, allopatric
species are, in general, younger than sympatric
ones, and if bindin divergence were accelerated
during speciation then slowed down, they should
show more differences in this molecule per unit
time than sympatric ones. The opposite is true
(Fig. 14.2).
The most credible hypothesis to date for differ-
ences in the rates of bindin evolution is that they
are caused by differences in the intensity of sex-
ual selection and sexual conflict. Using variation in
bindin genotypes of females as a proxy for varia-
tion in the bindin receptor (with which bindin is
expected to show linkage disequilibrium), Palumbi
(1999) has found that sexual selection exists in Echi-
nometra mathaei. Eggs are fertilized at higher rates
by sperm carrying the same bindin allele. Using the
same proxy, Levitan and Farrell (2006) and Levi-
tan and Stapper (2010) showed in Strongylocentrotus
franciscanus and S. purpuratus that sperm density
and the danger of polyspermy establish different
selective regimes for various bindin alleles. At low
sperm densities, most offspring are produced by
the union of sperm and egg possessing bindin
alleles that are most common in the population.
At high sperm densities, rare alleles leave behind
the most offspring, because common alleles, caus-
ing fast fertilization, result in polyspermic zygotes,
which fail to develop. Thus, there is always selec-
tion on males to effect fast fertilization, but females
in high sperm densities benefit from having alle-
les that retard fertilization: a typical sexual conflict
situation. Depending on ecological conditions, sex-
ual conflict can occur in some populations but not
others, thus resulting in different rates of bindin
evolution.
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
RATES OF SEA URCHIN BINDIN EVOLUTION 141
14.5 Conclusions and future prospects
In comparison to other invertebrate reproductive
proteins, bindin evolves moderately rapidly in
some genera and slowly in others. Selective reasons
for the differences that cause these dissimilarities in
rates are still the subject of speculation, but they
may well be related to fertilization environments
and intraspecific processes. Interspecific processes,
such as reinforcement, can also not be ruled out.
There may well be no universal explanation for the
presence or absence of positive selection in different
sea urchin taxa. Gametic proteins are often brought
up as examples of rapid evolution. Fast evolution is
certainly true for each of these proteins in the partic-
ular genus in which they have been studied. How-
ever, in a great many of the documented cases of
fast molecular evolution, the evidence comes only
from a small fraction of taxa. Data on sea urchin
bindin, though far from covering the entire echinoid
class, derive from multiple genera. This broader
taxonomic coverage alone may explain why more
diversity in the mode of evolution of this molecule
has been documented than has been found in other
invertebrate reproductive proteins.
Future laboratory studies linking the structure
of different bindin alleles with the specificity of
fertilization would be of great benefit in under-
standing the evolution of this molecule. We already
know which amino acids evolve under selection,
but we will need to determine the functional rea-
sons for such selection. Additional understanding
of the sources of natural selection on this molecule
and the rate of its evolution would come from com-
parative studies that link fertilization ecology in
nature with the success of particular bindin alle-
les. Simply characterizing species as sympatric or
allopatric on the basis of their geographic distri-
bution is not adequate for determining the role
of reinforcement or other interspecific processes in
bindin evolution. Ultimately, interest in the evolu-
tion of bindin and similar molecules stems from
our desire to understand the process of speciation
and the role of sexual selection in the evolution
of reproductive isolation. In that respect, assessing
the importance of bindin as a reproductive isolation
barrier between species relies on studies that are
not aimed directly at this molecule alone. Whether
bindin is involved in speciation depends not just
on the species-specificity of its interactions with its
receptor but on the probability that gametes of two
closely related sea urchin species will encounter
each other in nature. Even when gametic interac-
tions are, in fact, species-specific, it is still neces-
sary to determine whether bindin or some other
molecule, acting earlier in the sequence of fertiliza-
tion, is responsible. Thus, information on habitat
separation, reproductive timing, and pre-spawning
chemical communication as well as on the role of
other reproductive molecules is important in under-
standing whether intra- or interspecific interactions
mold the evolution of the bindin. Most of all, we
will need to link variation of bindin to variation
in its egg receptor. The study of EBR1 has been
retarded by its enormous size. Recent advances
in techniques for massive DNA sequencing have
made it practical to gather data on individual vari-
ation in large stretches of genetic material, and will
no doubt soon be applied to this problem.
Acknowledgments
We thank Laura Geyer and Santosh Jagadeeshan for
comments on the manuscript.
References
Biermann, C.H. (1998) The molecular evolution of sperm
bindin in six species of sea urchins (Echinoida: Strongy-
locentrotidae). Mol Biol Evol 15: 1761?71.
Calderon, I., Turon, X., and Lessios, H.A. (2009) Character-
ization of the sperm molecule bindin in the sea urchin
genus Paracentrotus. J Mol Evol 68: 366?76.
Calderon, I., Ventura, C.R.R., Turon, X., and Lessios,
H.A. (2010) Genetic divergence and assortative mating
between colour morphs of the sea urchin Paracentrotus
gaimardi. Mol Ecol 19: 484?93.
Civetta, A. and Singh, R.S. (1998) Sex-related genes, direc-
tional sexual selection, and speciation. Mol Biol Evol 15:
901?9.
Dunn, K.A., Bielawski, J.P., and Yang, Z. (2001) Substitu-
tion rates in Drosophila nuclear genes: implications for
translational selection. Genetics 157: 295?305.
Gao, B., Klein, L.E., Britten, R.J., and Davidson, E.H. (1986)
Sequence of mRNA coding for bindin, a species-specific
sea urchin sperm protein required for fertilization. Proc
Natl Acad Sci U S A 83: 8634?8.
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
142 RAPIDLY EVOLVING GENES AND GENETIC SYSTEMS
Geyer, L.B. and Palumbi, S.R. (2003) Reproductive charac-
ter displacement and the genetics of gamete recognition
in tropical sea urchins. Evolution 57, 1049?60.
Geyer, L.B. and Lessios, H.A. (2009) Lack of character dis-
placement in the male recognition molecule, bindin, in
Altantic sea urchins of the genus Echinometra. Mol Biol
Evol 26: 2135?46.
Hellberg, M.E. and Vacquier, V,D. (1999) Rapid evolution
of fertilization selectivity and lysin cDNA sequences in
teguline gastropods. Mol Biol Evol 16: 839?48.
Hellberg, M.E., Moy, G.W., and Vacquier, V.D. (2000) Pos-
itive selection and propeptide repeats promote rapid
interspecific divergence of a gastropod sperm protein.
Mol Biol Evol 17: 458?66.
Jordan, D. S. (1905) The origin of species through isolation.
Science 22: 545?62.
Kamei, N. and Glabe, C.G. (2003) The species-specific egg
receptor for sea urchin sperm adhesion is EBR1, a novel
ADAMTS protein. Genes Dev 17: 2502?7.
Kumar, S., Filipski, A., Swarna, V., Walker, A., and Hedges,
S.B. (2005) Placing confidence limits on the molecular
age of the human-chimpanzee divergence. Proc Natl
Acad Sci U S A 102: 18842?7.
Lessios, H.A. (2007) Reproductive isolation between
species of sea urchins. Bull Mar Sci 81: 191?208.
Lessios, H.A. (2008) The Great American Schism: Diver-
gence of marine organisms after the rise of the Cen-
tral American Isthmus. Annu Rev Ecol Evol Systema 39:
63?91.
Lessios, H.A. (2010) Speciation in sea urchins. In L.G. Har-
ris, S.A. B?ttger, C.W. Walker, and M.P. Lesser (Eds)
Echinoderms: Durham. Proceedings of the 12th Echinoderm
Conference, Durham, New Hampshire, pp. 91?101. Lon-
don: CRC Press.
Lessios, H.A. (2011) Speciation genes in free-spawning
marine invertebrates. Integr Comp Biol 51(3): 456?65.
Levitan, D.R. and Ferrell, D.L. (2006) Selection on gamete
recognition proteins depends on sex, density, and geno-
type frequency. Science 312: 267?9.
Levitan, D.R. and Stapper, A.P. (2010) Simultaneous pos-
itive and negative frequency-dependent selection on
sperm bindin, a gamete recognition protein in the
sea urchin Strongylocentrotus purpuratus. Evolution 64:
785?97.
Lopez, A., Miraglia, S.J., and Glabe, C.G. (1993) Struc-
ture/function analysis of the sea-urchin sperm adhesive
protein bindin. Dev Biol 156: 24?33.
McCartney, M.A. and Lessios, H.A. (2004) Adaptive evo-
lution of sperm bindin tracks egg incompatibility in
neotropical sea urchins of the genus Echinometra. Mol
Biol Evol 21: 732?45.
Metz, E.C. and Palumbi, S.R. (1996) Positive selection and
sequence rearrangements generate extensive polymor-
phism in the gamete recognition protein bindin. Mol Biol
Evol 13: 397?406.
Metz, E.C., Gomez-Gutierrez, G., and Vacquier, V.D.
(1998a) Mitochondrial DNA and bindin gene sequence
evolution among allopatric species of the sea urchin
genus Arbacia. Mol Biol Evol 15: 185?95.
Metz, E.C., Robles-Sikisaka, R., and Vacquier, V.D. (1998b)
Nonsynonymous substitution in abalone sperm fertil-
ization genes exceeds substitution in introns and mito-
chondrial DNA. Proc Natl Acad Sci U S A 95: 10676?81.
Palumbi, S.R. (1999) All males are not created equal: fer-
tility differences depend on gamete recognition poly-
morphisms in sea urchins. Proc Natl Acad Sci U S A 96:
12632?7.
Palumbi, S.R. (2009) Speciation and the evolution of
gamete recognition genes: Pattern and process. Heredity
102: 66?76.
Papadopoulou, A., Anastasiou, I., and Vogler, A.P. (2010)
Revisiting the insect mitochondrial molecular clock:
The mid-Aegean Trench calibration. Mol Biol Evol 27:
1659?72.
Patino, S., Aagaard, J.E., MacCoss, M.J., Swanson, W.J.,
and Hart, M.W. (2009) Bindin from a sea star. Evol Dev
11: 376?81.
Rocha, S., Lucio, M., Pereira, M.C., Reis, S., and Brezesin-
ski, G. (2008) The conformation of fusogenic B18 pep-
tide in surfactant solutions. J Peptide Sci 14: 436?41.
Swanson, W.J. and Vacquier, V.D. (2002a) The rapid evolu-
tion of reproductive proteins. Nat Rev Genet 3: 137?44.
Swanson, W.J. and Vacquier, V.D. (2002b) Reproductive
protein evolution. Annu Rev Ecol Systemat 33: 161?79.
Tsaur, S.-C. and Wu, C.-I. (1997) Positive selection and
the molecular evolution of a gene of male reproduction,
Acp26Aa of Drosophila. Mol Biol Evol 14: 544?9.
Vacquier, V.D. and Moy, G.W. (1977) Isolation of bindin:
The protein responsible for adhesion of sperm to sea
urchin eggs. Proc Natl Acad Sci U S A 74: 2456?60.
Vacquier, V.D. and Swanson, W.J. (2011) Selection in
the rapid evolution of gamete recognition proteins in
marine invertebrates. Cold Spring Harb Perspect Biol 3:
a002931.
Vacquier, V.D., Swanson, W.J., and Hellberg, M.E. (1995)
What have we learned about sea urchin sperm bindin?
Dev Growth Differ 37: 1?10.
Vieira, A. and Miller, D.J. (2006) Gamete interaction: Is it
species-specific? Mol Reprod Dev 73: 1422?9.
Wyckoff, G.J., Wang, W., and Wu, C.I. (2000) Rapid evolu-
tion of male reproductive genes in the descent of man.
Nature 403: 304?8.
OUP CORRECTED PROOF ? FINAL, 24/5/2012, SPi
RATES OF SEA URCHIN BINDIN EVOLUTION 143
Zigler, K.S. (2008) The evolution of sea urchin sperm
bindin. Int J Dev Biol 52: 791?6.
Zigler, K.S. and Lessios, H.A. (2003a) 250 million years of
bindin evolution. Biol Bull 205: 8?15.
Zigler, K.S. and Lessios, H.A. (2003b) Evolution of bindin
in the pantropical sea urchin Tripneustes: Compar-
isons to bindin of other genera. Mol Biol Evol 20:
220?31.
Zigler, K.S. and Lessios, H.A. (2004) Speciation on the
coasts of the new world: Phylogeography and the evo-
lution of bindin in the sea urchin genus Lytechinus.
Evolution 58: 1225?41.
Zigler, K.S., Raff, E.C., Popodi, E., Raff, R.A., and Lessios,
H.A. (2003) Adaptive evolution of bindin in the genus
Heliocidaris is correlated with the shift to direct develop-
ment. Evolution 57: 2293?302.
Zigler, K.S., Byrne, M., Raff, E.C., Lessios, H.A., and Raff,
R.A. (in press) Natural hybridization in the sea urchin
genus Pseudoboletia between species without apparent
barriers to gamete recognition. Evolution.