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Genomic and functional evolution of the Drosophila melanogaster sperm proteome

Abstract

In addition to delivering a haploid genome to the egg, sperm have additional critical functions, including egg activation, origination of the zygote centrosome and delivery of paternal factors1,2. Despite this, existing knowledge of the molecular basis of sperm form and function is limited. We used whole-sperm mass spectrometry to identify 381 proteins of the Drosophila melanogaster sperm proteome (DmSP). This approach identified mitochondrial, metabolic and cytoskeletal proteins, in addition to several new functional categories. We also observed nonrandom genomic clustering of sperm genes and underrepresentation on the X chromosome. Identification of widespread functional constraint on the proteome indicates that sexual selection has had a limited role in the overall evolution of D. melanogaster sperm. The relevance of the DmSP to the study of mammalian sperm function and fertilization mechanisms is demonstrated by the identification of substantial homology between the DmSP and proteins of the mouse axoneme accessory structure.

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Figure 1: Drosophila melanogaster sperm purification and 2D gel electrophoresis.
Figure 2: Molecular functional composition of the DmSP based on Gene Ontology annotation.
Figure 3: Genome distribution of the DmSP.
Figure 4: Evolutionary analysis of the DmSP.

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Acknowledgements

We thank S. Pitnick for assistance with sperm isolation and purification and C. Bergman, L. Hurst, J. Fay and B. Heath for discussions and constructive comments concerning the manuscript. We would also like to thank Z.N. Freeman, M. Karr, J. MacNaughton and D. Knowles for technical support. We also acknowledge the expertise of R. Reed at the University of California San Francisco protein sequencing facility. This work was funded in part by a Wolfson-Royal Society Merit Award, the Biotechnology and Biosciences Research Council (BBC0076701) and the US National Science Foundation (MCB-0135166) to T.L.K. and a Ruth L. Kirschstein National Research Service Award to S.D.

Author information

Authors and Affiliations

Authors

Contributions

This study was conceived and initiated by T.L.K. at the University of Chicago; protein identification was performed by S.A.B., J.S., D.F.H and U.G.; bioinformatic and evolutionary analyses were performed by S.D. and T.L.K.; S.D. and T.L.K. contributed to the writing of the paper.

Corresponding author

Correspondence to Timothy L Karr.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

ESI-MS MS spectra of a single hit assignment and deduced amino acid sequence for D. melanogaster gene CG31644. (PDF 208 kb)

Supplementary Table 1

Genes encoding the Drosophila melanogaster sperm proteome. (PDF 268 kb)

Supplementary Table 2

Peptides identified by mass spectrometry. (PDF 361 kb)

Supplementary Table 3

Membrane, signaling and neural genes of the DmSP. (PDF 78 kb)

Supplementary Table 4

2D gel/MALDI-TOF identification of D. melanogaster sperm proteome. (PDF 61 kb)

Supplementary Table 5

Homologous mouse sperm flagellum accessory structure proteins. (PDF 67 kb)

Supplementary Table 6

DmSP evolutionary rates. (PDF 75 kb)

Supplementary Table 7

10% of DmSP genes with the highest Ka/Ks ratios. (PDF 25 kb)

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Dorus, S., Busby, S., Gerike, U. et al. Genomic and functional evolution of the Drosophila melanogaster sperm proteome. Nat Genet 38, 1440–1445 (2006). https://doi.org/10.1038/ng1915

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