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CACN‐1 and its Interacting Proteins are Important Regulators of Somatic Development in Caenorhabditis elegans
Author(s) -
Hugunin Julie
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.600.2
Subject(s) - germline , biology , caenorhabditis elegans , gonad , somatic cell , microbiology and biotechnology , genetics , phenotype , sperm , gene , anatomy
With its easily visible U‐shaped gonad arm, Caenorhabditis elegans provides an excellent model to study germline development. This hermaphroditic nematode species has developed a mechanism to regulate the differentiation of its germline stem cells throughout development. C. elegans produce sperm until the onset of adulthood, when the worm permanently switches to producing oocytes. The Cram lab has identified the highly conserved protein CACN‐1 as an important player in proper germline differentiation. Loss of CACN‐1 leads to a masculinization of the germline where gonad arms are either exclusively filled with sperm, or contain a few irregular oocytes in addition to sperm. Previously, the lab identified CACN‐1 interacting proteins using tandem affinity purification and proteomics (TAP) 1 . CACN‐1 interactors grouped into a novel network suggest that CACN‐1 interacts with the spliceosome 1 . Depletion of some of these interactors in wild type animals also caused germline defects in differentiation and development comparable to CACN‐1. DAPI, a nuclear stain, revealed that gonad arms that lacked CACN‐1 and many of its interactors exhibited endomitotically duplicating oocytes (Emo phenotype) in a significant number of gonad arms containing abnormal oocytes. Due to the link between the Emo phenotype and ablated sheath cells 2 an actin stain was used to examine sheath cell presence and germline phenotypes when these proteins were depleted via RNAi. As expected depletion of CACN‐1 and its interacting proteins lead to defects in sheath actin staining that correlated with Emo and sperm containing gonad arms. In addition, loss of these proteins in a worm strain that is only susceptible to RNAi in the somatic cells caused germline defects comparable to those observed in wildtype animals. Our data suggests that this novel network of proteins plays an essential role in somatic gonad development that has a direct effect on germline differentiation and maintenance. Ultimately a better understanding of CACN‐1 and its network of proteins in C.elegans may elucidate the role of splicing and translational regulation of gene expression in other processes, including disease and development, across species. Support or Funding Information A special thanks is extended to Alyssa Cecchetelli, Michael Doherty and Erin Cram, as well as the entire Cram Lab, Northeastern University's Biology Department for supporting this research, and the NIH (R01 GM85077) for funding.