The regulation of SIRT2 function by cyclin-dependent kinases affects cell motility.
Cyclin-dependent kinases (Cdks) fulfill key functions in many cellular processes, including cell cycle progression and cytoskeletal dynamics. A limited number of Cdk substrates have been identified with few demonstrated to be regulated by Cdk-dependent phosphorylation. We identify on protein expression arrays novel cyclin E-Cdk2 substrates, including SIRT2, a member of the Sirtuin family of NAD(+)-dependent deacetylases that targets alpha-tubulin. We define Ser-331 as the site phosphorylated by cyclin E-Cdk2, cyclin A-Cdk2, and p35-Cdk5 both in vitro and in cells. Importantly, phosphorylation at Ser-331 inhibits the catalytic activity of SIRT2. Gain- and loss-of-function studies demonstrate that SIRT2 interfered with cell adhesion and cell migration. In postmitotic hippocampal neurons, neurite outgrowth and growth cone collapse are inhibited by SIRT2. The effects provoked by SIRT2, but not those of a nonphosphorylatable mutant, are antagonized by Cdk-dependent phosphorylation. Collectively, our findings identify a posttranslational mechanism that controls SIRT2 function, and they provide evidence for a novel regulatory circuitry involving Cdks, SIRT2, and microtubules.
Pandithage
Ruwin
R
Abteilung Biochemie und Molekularbiologie, Institut für Biochemie, Universitätsklinikum, Rheinisch-Westfälische Technische Hochschule Aachen University, 52057 Aachen, Germany.
Lilischkis
Richard
R
Harting
Kai
K
Wolf
Alexandra
A
Jedamzik
Britta
B
Lüscher-Firzlaff
Juliane
J
Vervoorts
Jörg
J
Lasonder
Edwin
E
Kremmer
Elisabeth
E
Knöll
Bernd
B
Lüscher
Bernhard
B
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002465:Cell Movement / Q000235:genetics*; D019925:Cyclin A / Q000235:genetics / Q000378:metabolism; D019927:Cyclin E / Q000235:genetics / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000235:genetics / Q000378:metabolism; D051360:Cyclin-Dependent Kinase 5 / Q000235:genetics / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism*; D020439:Growth Cones / Q000378:metabolism / Q000648:ultrastructure; D006367:HeLa Cells; D006624:Hippocampus / Q000196:embryology / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D051379:Mice; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational / Q000235:genetics; D012694:Serine / Q000378:metabolism; D056565:Sirtuin 2; D037761:Sirtuins / Q000235:genetics / Q000378:metabolism*
C475820:CCNE1 protein, human; D019925:Cyclin A; D019927:Cyclin E; D009419:Nerve Tissue Proteins; D015513:Oncogene Proteins; C089495:neuronal Cdk5 activator (p25-p35); D012694:Serine; D051360:Cyclin-Dependent Kinase 5; C495894:CDK2 protein, human; C495905:CDK5 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; C474199:SIRT2 protein, human; D056565:Sirtuin 2; D037761:Sirtuins