Role for human SIRT2 NAD-dependent deacetylase activity in control of mitotic exit in the cell cycle.
Studies of yeast have shown that the SIR2 gene family is involved in chromatin structure, transcriptional silencing, DNA repair, and control of cellular life span. Our functional studies of human SIRT2, a homolog of the product of the yeast SIR2 gene, indicate that it plays a role in mitosis. The SIRT2 protein is a NAD-dependent deacetylase (NDAC), the abundance of which increases dramatically during mitosis and is multiply phosphorylated at the G(2)/M transition of the cell cycle. Cells stably overexpressing the wild-type SIRT2 but not missense mutants lacking NDAC activity show a marked prolongation of the mitotic phase of the cell cycle. Overexpression of the protein phosphatase CDC14B, but not its close homolog CDC14A, results in dephosphorylation of SIRT2 with a subsequent decrease in the abundance of SIRT2 protein. A CDC14B mutant defective in catalyzing dephosphorylation fails to change the phosphorylation status or abundance of SIRT2 protein. Addition of 26S proteasome inhibitors to human cells increases the abundance of SIRT2 protein, indicating that SIRT2 is targeted for degradation by the 26S proteasome. Our data suggest that human SIRT2 is part of a phosphorylation cascade in which SIRT2 is phosphorylated late in G(2), during M, and into the period of cytokinesis. CDC14B may provoke exit from mitosis coincident with the loss of SIRT2 via ubiquitination and subsequent degradation by the 26S proteasome.
Dryden
Sylvia C
SC
Program in Molecular Biology and Genetics, Karmanos Cancer Institute and Wayne State University, Detroit, Michigan 48201, USA.
Nahhas
Fatimah A
FA
Nowak
James E
JE
Goustin
Anton-Scott
AS
Tainsky
Michael A
MA
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000918:Antibody Specificity; D002453:Cell Cycle / Q000502:physiology*; D002478:Cells, Cultured; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D054637:Dual-Specificity Phosphatases; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008938:Mitosis; D016297:Mutagenesis, Site-Directed; D009243:NAD / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000187:drug effects / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D046988:Proteasome Endopeptidase Complex; D020033:Protein Isoforms; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D056565:Sirtuin 2; D037761:Sirtuins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014162:Transfection; D025801:Ubiquitin / Q000378:metabolism
D015853:Cysteine Proteinase Inhibitors; D009842:Oligopeptides; D020033:Protein Isoforms; D025801:Ubiquitin; D009243:NAD; C067713:lactacystin; D010744:Phosphoric Monoester Hydrolases; C120902:CDC14A protein, human; C474200:CDC14B protein, human; D054637:Dual-Specificity Phosphatases; D017027:Protein Tyrosine Phosphatases; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; C474199:SIRT2 protein, human; D056565:Sirtuin 2; D037761:Sirtuins; D000111:Acetylcysteine; C078846:epoxomicin