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Turner BE, Basecke SM, Bazan GC, et al. Proteomic identification of germline proteins in Caenorhabditis elegans. Worm. 2015;4(1):e1008903doi: 10.1080/21624054.2015.1008903.
Turner, B. E., Basecke, S. M., Bazan, G. C., Dodge, E. S., Haire, C. M., Heussman, D. J., Johnson, C. L., Mukai, C. K., Naccarati, A. M., Norton, S. J., Sato, J. R., Talavera, C. O., Wade, M. V., & Hillers, K. J. (2015). Proteomic identification of germline proteins in Caenorhabditis elegans. Worm, 4(1), e1008903. https://doi.org/10.1080/21624054.2015.1008903
Turner, B Elizabeth, et al. "Proteomic identification of germline proteins in Caenorhabditis elegans." Worm vol. 4,1 (2015): e1008903. doi: https://doi.org/10.1080/21624054.2015.1008903
Turner BE, Basecke SM, Bazan GC, Dodge ES, Haire CM, Heussman DJ, Johnson CL, Mukai CK, Naccarati AM, Norton SJ, Sato JR, Talavera CO, Wade MV, Hillers KJ. Proteomic identification of germline proteins in Caenorhabditis elegans. Worm. 2015 Feb 09;4(1):e1008903. doi: 10.1080/21624054.2015.1008903. eCollection 2015. PMID: 26435885; PMCID: PMC4590026.
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Worm. 2015 Feb 09;4(1):e1008903. doi: 10.1080/21624054.2015.1008903. eCollection 2015.

Proteomic identification of germline proteins in Caenorhabditis elegans.

Worm

B Elizabeth Turner, Sophia M Basecke, Grace C Bazan, Eric S Dodge, Cassy M Haire, Dylan J Heussman, Chelsey L Johnson, Chelsea K Mukai, Adrianna M Naccarati, Sunny-June Norton, Jennifer R Sato, Chihara O Talavera, Michael V Wade, Kenneth J Hillers

Affiliations

  1. 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.
  2. Department of Biological Sciences; California Polytechnic State University ; San Luis Obispo, CA USA.
  3. Department of Chemistry and Biochemistry; California Polytechnic State University ; San Luis Obispo, CA USA ;

PMID: 26435885 PMCID: PMC4590026 DOI: 10.1080/21624054.2015.1008903

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.

Keywords: 2D gel electrophoresis; RNA interference; comparative proteomics; fertility; gametogenesis; meiosis

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