title: Proteomic identification of germline proteins in Caenorhabditis elegans.
abstract: Sexual reproduction involves fusion of 2 haploid gametes to form diploid offspring with genetic contributions from both parents. Gamete formation represents a unique developmental program involving the action of numerous germline-specific proteins. In an attempt to identify novel proteins involved in reproduction and embryonic development, we have carried out a proteomic characterization of the process in Caenorhabditis elegans. To identify candidate proteins, we used 2D gel electrophoresis (2DGE) to compare protein abundance in nucleus-enriched extracts from wild-type C. elegans, and in extracts from mutant worms with greatly reduced gonads (glp-4(bn2) worms reared at 25°C); 84 proteins whose abundance correlated with germline presence were identified. To validate candidates, we used feeding RNAi to deplete candidate proteins, and looked for reduction in fertility and/or germline cytological defects. Of 20 candidates so screened for involvement in fertility, depletion of 13 (65%) caused a significant reduction in fertility, and 6 (30%) resulted in sterility (<5 % of wild-type fertility). Five of the 13 proteins with demonstrated roles in fertility have not previously been implicated in germline function. The high frequency of defects observed after RNAi depletion of candidate proteins suggests that this approach is effective at identifying germline proteins, thus contributing to our understanding of this complex organ.
authors.affiliation: Department of Chemistry and Biochemistry; California Polytechnic State University ; San Luis Obispo, CA USA ; ; Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: B Elizabeth
authors.initials: BE
authors.lastname: Turner
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Sophia M
authors.initials: SM
authors.lastname: Basecke
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Grace C
authors.initials: GC
authors.lastname: Bazan
authors.affiliation: Department of Chemistry and Biochemistry; California Polytechnic State University ; San Luis Obispo, CA USA ;
authors.forename: Eric S
authors.initials: ES
authors.lastname: Dodge
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Cassy M
authors.initials: CM
authors.lastname: Haire
authors.affiliation: Department of Chemistry and Biochemistry; California Polytechnic State University ; San Luis Obispo, CA USA ;
authors.forename: Dylan J
authors.initials: DJ
authors.lastname: Heussman
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Chelsey L
authors.initials: CL
authors.lastname: Johnson
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Chelsea K
authors.initials: CK
authors.lastname: Mukai
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Adrianna M
authors.initials: AM
authors.lastname: Naccarati
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Sunny-June
authors.initials: SJ
authors.lastname: Norton
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Sato
authors.affiliation: Department of Chemistry and Biochemistry; California Polytechnic State University ; San Luis Obispo, CA USA ;
authors.forename: Chihara O
authors.initials: CO
authors.lastname: Talavera
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Michael V
authors.initials: MV
authors.lastname: Wade
authors.affiliation: Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
authors.forename: Kenneth J
authors.initials: KJ
authors.lastname: Hillers
keywords: 2D gel electrophoresis; RNA interference; comparative proteomics; fertility; gametogenesis; meiosis