Saccharomyces CDK1 phosphorylates Rad53 kinase in metaphase, influencing cellular morphogenesis.
Rad53 is an essential protein kinase governing DNA damage and replication stress checkpoints in budding yeast. It also appears to be involved in cellular morphogenesis processes. Mass spectrometry analyses revealed that Rad53 is phosphorylated at multiple SQ/TQ and at SP/TP residues, which are typical consensus sites for phosphatidylinositol 3-kinase-related kinases and CDKs, respectively. Here we show that Clb-CDK1 phosphorylates Rad53 at Ser(774) in metaphase. This phosphorylation event does not influence the DNA damage and replication checkpoint roles of Rad53, and it is independent of the spindle assembly checkpoint network. Moreover, the Ser-to-Asp mutation, mimicking a constitutive phosphorylation state at site 774, causes sensitivity to calcofluor, supporting a functional linkage between Rad53 and cellular morphogenesis.
Diani
Laura
L
Dipartimento di Scienze Biomolecolari e Biotecnologie, Universita' degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy.
Colombelli
Claudia
C
Nachimuthu
Benjamin Tamilselvan
BT
Donnianni
Roberto
R
Plevani
Paolo
P
Muzi-Falconi
Marco
M
Pellicioli
Achille
A
D000483:Alleles; D001224:Aspartic Acid / Q000737:chemistry; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002469:Cell Separation; D064447:Checkpoint Kinase 2; D004249:DNA Damage; D008954:Models, Biological; D016296:Mutagenesis; D009154:Mutation; D015739:Nocodazole / Q000494:pharmacology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D012694:Serine / Q000737:chemistry
D018797:Cell Cycle Proteins; D029701:Saccharomyces cerevisiae Proteins; D001224:Aspartic Acid; D012694:Serine; D064447:Checkpoint Kinase 2; D017346:Protein Serine-Threonine Kinases; D016203:CDC2 Protein Kinase; C067022:RAD53 protein, S cerevisiae; D015739:Nocodazole