Phosphorylation of cyclin dependent kinase 4 on tyrosine 17 is mediated by Src family kinases.
Cyclin dependent kinase 4 is a key regulator of the cell cycle and its activity is frequently deregulated in cancer. The activity of cyclin dependent kinase 4 is controlled by multiple mechanisms, including phosphorylation of tyrosine 17. This site is equivalent to tyrosine 15 of cyclin dependent kinase 1, which undergoes inhibitory phosphorylation by WEE1 and MYT1; however, the kinases that phosphorylate cyclin dependent kinase 4 on tyrosine 17 are still unknown. In the present study, we generated a phosphospecific antibody to the tyrosine 17-phosphorylated form of cyclin dependent kinase 4, and showed that this site is phosphorylated to a low level in asynchronously proliferating HCT116 cells. We purified tyrosine 17 kinases from HeLa cells and found that the Src family non-receptor tyrosine kinase C-YES contributes a large fraction of the tyrosine 17 kinase activity in HeLa lysates. C-YES also phosphorylated cyclin dependent kinase 4 when transfected into HCT116 cells, and treatment of cells with Src family kinase inhibitors blocked the tyrosine 17 phosphorylation of cyclin dependent kinase 4. Taken together, the results obtained in the present study provide the first evidence that Src family kinases, but not WEE1 or MYT1, phosphorylate cyclin dependent kinase 4 on tyrosine 17, and help to resolve how the phosphorylation of this site is regulated.
Martin
Nicholas G
NG
Centre for Cancer Therapeutics at The Institute of Cancer Research, Cancer Research UK, Haddow Laboratories, 15 Cotswold Road, Sutton, Surrey, UK.
McAndrew
Peter C
PC
Eve
Paul D
PD
Garrett
Michelle D
MD
D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000378:metabolism; D051358:Cyclin-Dependent Kinase 4 / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051575:Proto-Oncogene Proteins c-yes / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014443:Tyrosine / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
D018797:Cell Cycle Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D051575:Proto-Oncogene Proteins c-yes; C499349:WEE1 protein, human; D019061:src-Family Kinases; C490679:PKMYT1 protein, human; D017346:Protein Serine-Threonine Kinases; D051358:Cyclin-Dependent Kinase 4