title: Mitotic exit kinase Dbf2 directly phosphorylates chitin synthase Chs2 to regulate cytokinesis in budding yeast.
abstract: How cell cycle machinery regulates extracellular matrix (ECM) remodeling during cytokinesis remains poorly understood. In the budding yeast Saccharomyces cerevisiae, the primary septum (PS), a functional equivalent of animal ECM, is synthesized during cytokinesis by the chitin synthase Chs2. Here, we report that Dbf2, a conserved mitotic exit kinase, localizes to the division site after Chs2 and directly phosphorylates Chs2 on several residues, including Ser-217. Both phosphodeficient (chs2-S217A) and phosphomimic (chs2-S217D) mutations cause defects in cytokinesis, suggesting that dynamic phosphorylation-dephosphorylation of Ser-217 is critical for Chs2 function. It is striking that Chs2-S217A constricts asymmetrically with the actomyosin ring (AMR), whereas Chs2-S217D displays little or no constriction and remains highly mobile at the division site. These data suggest that Chs2 phosphorylation by Dbf2 triggers its dissociation from the AMR during the late stage of cytokinesis. Of interest, both chs2-S217A and chs2-S217D mutants are robustly suppressed by increased dosage of Cyk3, a cytokinesis protein that displays Dbf2-dependent localization and also stimulates Chs2-mediated chitin synthesis. Thus Dbf2 regulates PS formation through at least two independent pathways: direct phosphorylation and Cyk3-mediated activation of Chs2. Our study establishes a mechanism for direct cell cycle control of ECM remodeling during cytokinesis.
authors.affiliation: Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: Younghoon
authors.initials: Y
authors.lastname: Oh
authors.forename: Kuang-Jung
authors.initials: KJ
authors.lastname: Chang
authors.forename: Peter
authors.initials: P
authors.lastname: Orlean
authors.forename: Carsten
authors.initials: C
authors.lastname: Wloka
authors.forename: Raymond
authors.initials: R
authors.lastname: Deshaies
authors.forename: Erfei
authors.initials: E
authors.lastname: Bi
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D002686:Chitin / Q000378:metabolism; D002687:Chitin Synthase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D048749:Cytokinesis; D036681:Fluorescence Recovery After Photobleaching; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism*; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000254:growth & development / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D059008:Time-Lapse Imaging
chemical_list: D018797:Cell Cycle Proteins; C418803:Cyk3 protein, S cerevisiae; D008869:Microtubule-Associated Proteins; D029701:Saccharomyces cerevisiae Proteins; D002686:Chitin; D002687:Chitin Synthase; C400821:chitin synthase 2; C085768:DBF2 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases