Rod1, an arrestin-related protein, is phosphorylated by Snf1-kinase in Saccharomyces cerevisiae.
Metazoan arrestin proteins bind to seven-transmembrane proteins, mediate their internalization and play central roles in the subsequent signal transduction pathway. In Saccharomyces cerevisiae, there are several arrestin-related proteins. One of those proteins, Rod1, has been identified to have the ability to confer resistance to o-dinitrobenzene. We found that Rod1 interacted with Snf4, a subunit of Snf1-kinase complex. Both snf4 and snf1 mutants were also sensitive to the drug and the kinase activity of Snf1 was required for the drug tolerance. In immunoblotting analysis, the Rod1 protein was phosphorylated in an Snf1-dependent manner in vivo, and the phosphorylation of the serine residue 447 of Rod1 was responsible for the band-shift. Furthermore, the Rod1 protein was directly phosphorylated by Snf1-kinase in vitro. The substitution of the serine residue 447 to alanine slightly enhanced the resistance to the drug. We discuss possible functions of Rod1.
Shinoda
Junro
J
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
Kikuchi
Yoshiko
Y
D055372:AMP-Activated Protein Kinases; D000409:Alanine / Q000235:genetics; D019943:Amino Acid Substitution; D019393:Arrestin / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D004136:Dinitrobenzenes / Q000494:pharmacology; D025141:Drug Resistance, Fungal; D005656:Fungal Proteins / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D008565:Membrane Proteins; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D012694:Serine / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism
D019393:Arrestin; D002352:Carrier Proteins; D004136:Dinitrobenzenes; D005656:Fungal Proteins; D008565:Membrane Proteins; C099318:ROD1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C040234:1,2-dinitrobenzene; D012694:Serine; C071570:SNF1-related protein kinases; D017346:Protein Serine-Threonine Kinases; C078651:SNF4 protein, S cerevisiae; D055372:AMP-Activated Protein Kinases; D000409:Alanine