Abstract
Tyrosine phosphorylation plays a central role in eukaryotic signal transduction. In yeast, MAP kinase pathways are regulated by tyrosine phosphorylation, and it has been speculated that other biochemical processes may also be regulated by tyrosine phosphorylation. Previous genetic and biochemical studies demonstrate that protein tyrosine phosphatases (PTPases) negatively regulate yeast MAP kinases. Here we report that deletion of PTP2 and PTP3 results in a sporulation defect, suggesting that tyrosine phosphorylation is involved in regulation of meiosis and sporulation. Deletion of PTP2 and PTP3 blocks cells at an early stage of sporulation before premeiotic DNA synthesis and induction of meiotic-specific genes. We observed that tyrosine phosphorylation of several proteins, including 52-, 43-, and 42-kDa proteins, was changed in ptp2Deltaptp3Delta homozygous deletion cells under sporulation conditions. The 42-kDa tyrosine-phosphorylated protein was identified as Mck1, which is a member of the GSK3 family of protein kinases and previously known to be phosphorylated on tyrosine. Mutation of MCK1 decreases sporulation efficiency, whereas mutation of RIM11, another GSK3 member, specifically abolishes sporulation; therefore, we investigated regulation of Rim11 by Tyr phosphorylation during sporulation. We demonstrated that Rim11 is phosphorylated on Tyr-199, and the Tyr phosphorylation is essential for its in vivo function, although Rim11 appears not to be directly regulated by Ptp2 and Ptp3. Biochemical characterizations indicate that tyrosine phosphorylation of Rim11 is essential for the activity of Rim11 to phosphorylate substrates. Our data demonstrate important roles of protein tyrosine phosphorylation in meiosis and sporulation
MeSH Terms
DNA, Fungal/biosynthesis
Fungal Proteins/chemistry,genetics,metabolism
Gene Deletion
Gene Expression
Genes, Essential/genetics,physiology
Genes, Fungal/genetics,physiology
Glycogen Synthase Kinase 3
Intracellular Signaling Peptides and Proteins
Meiosis/genetics
Molecular Weight
Phosphorylation
Phosphotyrosine/metabolism
Protein Serine-Threonine Kinases/genetics,metabolism
Protein Tyrosine Phosphatases/genetics,metabolism
Protein-Tyrosine Kinases/chemistry,genetics,metabolism
Saccharomyces cerevisiae/cytology,enzymology,genetics,physiology
Saccharomyces cerevisiae Proteins
Spores, Fungal/cytology,enzymology,genetics,physiology
Tyrosine/genetics,metabolism
Chemicals
DNA, Fungal
Fungal Proteins
Intracellular Signaling Peptides and Proteins
Saccharomyces cerevisiae Proteins
Phosphotyrosine
Tyrosine
Protein-Tyrosine Kinases
Protein Serine-Threonine Kinases
RIM11 protein, S cerevisiae
Glycogen Synthase Kinase 3
MCK1 protein, S cerevisiae
Protein Tyrosine Phosphatases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Zhan X L
Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
Hong Y
Zhu T
Mitchell A P
Deschenes R J
Guan K L
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