Proteomic analysis of in vivo 14-3-3 interactions in the yeast Saccharomyces cerevisiae.
The yeast Saccharomyces cerevisiae produces two 14-3-3 proteins, Bmh1 and Bmh2, whose exact functions have remained unclear. Here, we performed a comprehensive proteomic analysis using multistep immunoaffinity purification and mass spectrometry and identified 271 yeast proteins that specifically bind to Bmh1 and -2 in a phosphorylation-dependent manner. The identified proteins have diverse biochemical functions and cellular roles, including cell signaling, metabolism, and cell cycle regulation. Importantly, there are a number of protein subsets that are involved in the regulation of yeast physiology through a variety of cell signaling pathways, including stress-induced transcription, cell division, and chitin synthesis at the cell wall. In fact, we found that a yeast mutant deficient in Bmh1 and -2 had defects in signal-dependent response of the MAPK (Hog1 and Mpk1) cascade and exhibited an abnormal accumulation of chitin at the bud neck. We propose that Bmh1 and -2 are common regulators of many cell signaling modules and pathways mediated by protein phosphorylation and regulate a variety of biological events by coordinately controlling the identified multiplex phosphoprotein components.
Kakiuchi
Kazue
K
Department of Chemistry, Graduate School of Science, Tokyo Metropolitan University, Hachioji, Tokyo 192-0397, Japan.
Yamauchi
Yoshio
Y
Taoka
Masato
M
Iwago
Maki
M
Fujita
Tomoko
T
Ito
Takashi
T
Song
Si-Young
SY
Sakai
Akira
A
Isobe
Toshiaki
T
Ichimura
Tohru
T
D048948:14-3-3 Proteins / Q000378:metabolism*; D002686:Chitin / Q000096:biosynthesis; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism* / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology
D048948:14-3-3 Proteins; C075852:BMH1 protein, S cerevisiae; C097164:BMH2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D002686:Chitin; C088589:HOG1 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases; C072724:SLT2 protein, S cerevisiae