Identification of a human cDNA encoding a kinase-defective cdk5 isoform.
The cyclin-dependent kinase 5 (Cdk5) catalytic subunit is expressed in both cycling and noncycling cells and is present in many tissues. Neuronal and muscle cells contain the highest amount of this protein. The p35 protein, which is expressed solely in the brain, activates Cdk5. Cdk5 activity is involved in terminal differentiation of neurons and muscle cells. We attempted to clone cdk5 by PCR from a human fetal brain cDNA library. Surprisingly, we amplified two forms of the cdk5 gene, the wild type and a cdk5 variant that lacks the complete kinase domain VI. The variant is also found in SH-SY-5Y neuroblastoma cells but not in T-cells, HeLa cells, the thymus, and placental tissue. The protein encoded by the cdk5 variant, the Cdk5 isoform (Cdk5i), purifies with p35 when coexpressed in insect cells. The activity associated with the heterodimer Cdk5i/p35 is found to be appreciably weaker than the wild-type Cdk5/p35 kinase. Moreover, Cdk5i/p35 cannot autophosphorylate its two subunits as with Cdk5/p35. Interestingly, kinase-defective Cdk5i can abolish the activity of wild-type Cdk5 when both are coexpressed with p35 in insect cells, suggesting that Cdk5i may have a function in regulating Cdk5 activity in human cells too.
Moorthamer
M
M
Oncology Research, Novartis Pharma AG, Basel, Switzerland.
Zumstein-Mecker
S
S
Stephan
C
C
Mittl
P
P
Chaudhuri
B
B
D000595:Amino Acid Sequence; D001921:Brain; D003001:Cloning, Molecular; D051360:Cyclin-Dependent Kinase 5; D018844:Cyclin-Dependent Kinases; D018076:DNA, Complementary / Q000302:isolation & purification*; D005333:Fetus; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000172:deficiency / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009447:Neuroblastoma; D010920:Placenta; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency* / Q000235:genetics*; D013601:T-Lymphocytes; D013950:Thymus Gland; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D007527:Isoenzymes; D051360:Cyclin-Dependent Kinase 5; D017346:Protein Serine-Threonine Kinases; C495905:CDK5 protein, human; D018844:Cyclin-Dependent Kinases