title: Proteomic analysis reveals CACN-1 is a component of the spliceosome in Caenorhabditis elegans.
abstract: Cell migration is essential for embryonic development and tissue formation in all animals. cacn-1 is a conserved gene of unknown molecular function identified in a genome-wide screen for genes that regulate distal tip cell migration in the nematode worm Caenorhabditis elegans. In this study we take a proteomics approach to understand CACN-1 function. To isolate CACN-1-interacting proteins, we used an in vivo tandem-affinity purification strategy. Tandem-affinity purification-tagged CACN-1 complexes were isolated from C. elegans lysate, analyzed by mass spectrometry, and characterized bioinformatically. Results suggest significant interaction of CACN-1 with the C. elegans spliceosome. All of the identified interactors were screened for distal tip cell migration phenotypes using RNAi. Depletion of many of these factors led to distal tip cell migration defects, particularly a failure to stop migrating, a phenotype commonly seen in cacn-1 deficient animals. The results of this screen identify eight novel regulators of cell migration and suggest CACN-1 may participate in a protein network dedicated to high-fidelity gonad development. The composition of proteins comprising the CACN-1 network suggests that this critical developmental module may exert its influence through alternative splicing or other post-transcriptional gene regulation.
authors.affiliation: Biology Department, Northeastern University, Boston, Massachusetts 02115.
authors.forename: Michael F
authors.initials: MF
authors.lastname: Doherty
authors.affiliation: Department of Cancer Biology and Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, Massachusetts 02215 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Adelmant
authors.affiliation: Biology Department, Northeastern University, Boston, Massachusetts 02115.
authors.forename: Alyssa D
authors.initials: AD
authors.lastname: Cecchetelli
authors.affiliation: Department of Cancer Biology and Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, Massachusetts 02215 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Jarrod A
authors.initials: JA
authors.lastname: Marto
authors.affiliation: Biology Department, Northeastern University, Boston, Massachusetts 02115 e.cram@neu.edu.
authors.forename: Erin J
authors.initials: EJ
authors.lastname: Cram
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism*; D002465:Cell Movement; D009687:Nuclear Proteins / Q000378:metabolism*; D060066:Protein Interaction Maps; D040901:Proteomics; D034622:RNA Interference; D017381:Spliceosomes / Q000378:metabolism*
chemical_list: C000595442:CACN-1 protein, C elegans; D029742:Caenorhabditis elegans Proteins; D009687:Nuclear Proteins
keywords: C. elegans; cell migration; proteomics; spliceosome