title: In Vivo Interaction Proteomics in Caenorhabditis elegans Embryos Provides New Insights into P Granule Dynamics.
abstract: Studying protein interactions in whole organisms is fundamental to understanding development. Here, we combine in vivo expressed GFP-tagged proteins with quantitative proteomics to identify protein-protein interactions of selected key proteins involved in early C. elegans embryogenesis. Co-affinity purification of interaction partners for eight bait proteins resulted in a pilot in vivo interaction map of proteins with a focus on early development. Our network reflects known biology and is highly enriched in functionally relevant interactions. To demonstrate the utility of the map, we looked for new regulators of P granule dynamics and found that GEI-12, a novel binding partner of the DYRK family kinase MBK-2, is a key regulator of P granule formation and germline maintenance. Our data corroborate a recently proposed model in which the phosphorylation state of GEI-12 controls P granule dynamics. In addition, we find that GEI-12 also induces granule formation in mammalian cells, suggesting a common regulatory mechanism in worms and humans. Our results show that in vivo interaction proteomics provides unique insights into animal development.
authors.affiliation: From the ‡Max Delbrück Center for Molecular Medicine, D-13092 Berlin, Germany; §Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003;
authors.forename: Jia-Xuan
authors.initials: JX
authors.lastname: Chen
authors.affiliation: §Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003; ¶New York University Abu Dhabi, Abu Dhabi, United Arab Emirates;
authors.forename: Patricia G
authors.initials: PG
authors.lastname: Cipriani
authors.affiliation: §Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003;
authors.forename: Desirea
authors.initials: D
authors.lastname: Mecenas
authors.affiliation: §Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003; ‖INSERM, U1104, 13288 Marseille, France;
authors.forename: Jolanta
authors.initials: J
authors.lastname: Polanowska
authors.affiliation: §Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003; ¶New York University Abu Dhabi, Abu Dhabi, United Arab Emirates;
authors.forename: Fabio
authors.initials: F
authors.lastname: Piano
authors.affiliation: §Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003; ¶New York University Abu Dhabi, Abu Dhabi, United Arab Emirates; matthias.selbach@mdc-berlin.de kcg1@nyu.edu.
authors.forename: Kristin C
authors.initials: KC
authors.lastname: Gunsalus
authors.affiliation: From the ‡Max Delbrück Center for Molecular Medicine, D-13092 Berlin, Germany; **Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany. matthias.selbach@mdc-berlin.de kcg1@nyu.edu.
authors.forename: Matthias
authors.initials: M
authors.lastname: Selbach
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000032:analysis* / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002846:Chromatography, Affinity; D003594:Cytoplasmic Granules / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D013058:Mass Spectrometry; D010766:Phosphorylation; D060066:Protein Interaction Maps; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D000097603:Dyrk Kinases
chemical_list: D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D000097603:Dyrk Kinases; C000620624:GEI-12 protein, C elegans; C478059:MBK-2 protein, C elegans