RRC ID 46690
著者 Wilson LD, Sackett JM, Mieczkowski BD, Richie AL, Thoemke K, Rumbley JN, Kroft TL.
タイトル Fertilization in C. elegans requires an intact C-terminal RING finger in sperm protein SPE-42.
ジャーナル BMC Dev Biol
Abstract BACKGROUND:The C. elegans sperm protein SPE-42, a membrane protein of unknown structure and molecular function, is required for fertilization. Sperm from worms with spe-42 mutations appear normal but are unable to fertilize eggs. Sequence analysis revealed the presence of 8 conserved cysteine residues in the C-terminal cytoplasmic domain of this protein suggesting these residues form a zinc-coordinating RING finger structure.
RESULTS:We made an in silico structural model of the SPE-42 RING finger domain based on primary sequence analysis and previously reported RING structures. To test the model, we created spe-42 transgenes coding for mutations in each of the 8 cysteine residues predicted to coordinate Zn++ ions in the RING finger motif. Transgenes were crossed into a spe-42 null background and protein function was measured by counting progeny. We found that all 8 cysteines are required for protein function. We also showed that sequence differences between the C-terminal 29 and 30 amino acids in C. elegans and C. briggsae SPE-42 following the RING finger domain are not responsible for the failure of the C. briggsae SPE-42 homolog to rescue C. elegans spe-42 mutants.
CONCLUSIONS:The results suggest that a bona fide RING domain is present at the C-terminus of the SPE-42 protein and that this motif is required for sperm-egg interactions during C. elegans fertilization. Our structural model of the RING domain provides a starting point for further structure-function analysis of this critical region of the protein. The C-terminal domain swap experiment suggests that the incompatibility between the C. elegans and C. briggsae SPE-42 proteins is caused by small amino acid differences outside the C-terminal domain.
巻・号 11
ページ 10
公開日 2011-2-23
DOI 10.1186/1471-213X-11-10
PII 1471-213X-11-10
PMID 21345212
PMC PMC3053230
MeSH Amino Acid Sequence Animals Base Sequence Caenorhabditis elegans / genetics Caenorhabditis elegans / physiology* Caenorhabditis elegans Proteins / chemistry* Caenorhabditis elegans Proteins / metabolism* Cysteine / chemistry Cysteine / metabolism Fertilization Membrane Proteins / chemistry* Membrane Proteins / metabolism* Models, Molecular Molecular Sequence Data Mutation Polymerase Chain Reaction Protein Interaction Domains and Motifs Protein Structure, Tertiary RING Finger Domains* / genetics Sequence Analysis, Protein Sperm-Ovum Interactions Structure-Activity Relationship Zinc / chemistry
IF 2.0
引用数 7
WOS 分野 DEVELOPMENTAL BIOLOGY
リソース情報
線虫 tm2421