title: Large-Scale Analysis of Kinase Signaling in Yeast Pseudohyphal Development Identifies Regulation of Ribonucleoprotein Granules.
abstract: Yeast pseudohyphal filamentation is a stress-responsive growth transition relevant to processes required for virulence in pathogenic fungi. Pseudohyphal growth is controlled through a regulatory network encompassing conserved MAPK (Ste20p, Ste11p, Ste7p, Kss1p, and Fus3p), protein kinase A (Tpk2p), Elm1p, and Snf1p kinase pathways; however, the scope of these pathways is not fully understood. Here, we implemented quantitative phosphoproteomics to identify each of these signaling networks, generating a kinase-dead mutant in filamentous S. cerevisiae and surveying for differential phosphorylation. By this approach, we identified 439 phosphoproteins dependent upon pseudohyphal growth kinases. We report novel phosphorylation sites in 543 peptides, including phosphorylated residues in Ras2p and Flo8p required for wild-type filamentous growth. Phosphoproteins in these kinase signaling networks were enriched for ribonucleoprotein (RNP) granule components, and we observe co-localization of Kss1p, Fus3p, Ste20p, and Tpk2p with the RNP component Igo1p. These kinases localize in puncta with GFP-visualized mRNA, and KSS1 is required for wild-type levels of mRNA localization in RNPs. Kss1p pathway activity is reduced in lsm1Δ/Δ and pat1Δ/Δ strains, and these genes encoding P-body proteins are epistatic to STE7. The P-body protein Dhh1p is also required for hyphal development in Candida albicans. Collectively, this study presents a wealth of data identifying the yeast phosphoproteome in pseudohyphal growth and regulatory interrelationships between pseudohyphal growth kinases and RNPs.
authors.affiliation: Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, Michigan, United States of America; Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: Christian A
authors.initials: CA
authors.lastname: Shively
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan, United States of America.
authors.forename: Hye Kyong
authors.initials: HK
authors.lastname: Kweon
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: Kaitlyn L
authors.initials: KL
authors.lastname: Norman
authors.affiliation: Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan, United States of America; Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, Michigan, United States of America.
authors.forename: Dattatreya
authors.initials: D
authors.lastname: Mellacheruvu
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Sheidy
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Dobry
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, Indiana, United States of America.
authors.forename: Ivan
authors.initials: I
authors.lastname: Sabath
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: Eric E P
authors.initials: EE
authors.lastname: Cosky
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, Indiana, United States of America.
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Tran
authors.affiliation: Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan, United States of America; Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, Michigan, United States of America.
authors.forename: Alexey
authors.initials: A
authors.lastname: Nesvizhskii
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan, United States of America; Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, Michigan, United States of America.
authors.forename: Philip C
authors.initials: PC
authors.lastname: Andrews
authors.affiliation: Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, Michigan, United States of America; Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: Anuj
authors.initials: A
authors.lastname: Kumar
mesh_terms: D002176:Candida albicans / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D025301:Hyphae / Q000235:genetics* / Q000254:growth & development; D010641:Phenotype; D010766:Phosphorylation; D010770:Phosphotransferases / Q000096:biosynthesis* / Q000235:genetics; D012261:Ribonucleoproteins / Q000096:biosynthesis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics; D015398:Signal Transduction
chemical_list: D012261:Ribonucleoproteins; D029701:Saccharomyces cerevisiae Proteins; D010770:Phosphotransferases