Int. J. Dev. Biol. 52: 759 - 768 (2008)

https://doi.org/10.1387/ijdb.082581ss

Vol 52, Issue 5-6

Special Issue: Fertilization

Oyster sperm bindin is a combinatorial fucose lectin with remarkable intra-species diversity

Published: 1 July 2008

Stevan A. Springer¹, Gary W. Moy², Daniel S. Friend³, Willie J. Swanson⁴ and Victor D. Vacquier²

¹Department of Biology, University of Washington, Seattle, Washington, ²Scripps Institution of Oceanography, University of California San Diego, La Jolla, California, ³Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts and ⁴Department of Genome Sciences, University of Washington, Seattle WA, USA

Abstract

Sperm of the oyster, Crassostrea gigas, have ring-shaped acrosomes that, after exocytosis, bind the sperm to the egg vitelline layer. Isolated acrosomal rings contain proteins of various sizes: 35-, 48-, 63-, 75- and 88-kDa. These proteins, called bindins, have identical 24-residue signal peptides and conserved 97-residue N-terminal sequences, and they differ in mass because of the presence of between 1 and 5 tandemly repeated 134-residue fucose-binding lectin (F-lectin) domains. Southern blots suggest that oyster bindin is a single copy gene, but F-lectin repeat number and sequence are variable within and between individuals. Eight residues in the F-lectin fucose-binding groove are subject to positive diversifying selection, indicating a history of adaptive evolution at the lectin's active site. There is one intron in the middle of each F-lectin repeat, and recombination in this intron creates many combinations of repeat halves. Alternative splicing creates many additional size and sequence variants of the repeat array. Males contain full-length bindin cDNAs of all 5 possible sizes, but only one or two protein mass forms exist in each individual. Sequence analysis indicates that recombination and alternate splicing create hundreds, possibly thousands, of different bindin sequences in C. gigas. The extreme within-species sequence variation in the F-lectin sequence of oyster bindin is a novel finding; most male gamete-recognition proteins are much less variable. In experimental conditions oyster eggs have poor polyspermy blocks, and bindin diversity could be an evolutionary response by sperm to match egg receptors that have diversified to avoid being fertilized by multiple sperm.

Keywords

sperm-egg interaction, gamete recognition, invertebrate fertilization, positive selection, molecular evolution, alternative splicing, recombination