Analysis of p16INK4a and its interaction with CDK4.
The interaction between cyclin-dependent kinase 4 (CDK4) and its inhibitor p16INK4a (p16) was studied by random mutagenesis and yeast two-hybrid system. The gene encoding p16 was mutagenized randomly and the amino acid changes that affect the binding of p16 to CDK4 were identified. Several amino acid residues were shown to be important for the binding and many of these changes occur at residues conserved in all known human p16 family proteins Most of the mutant p16 proteins that failed to bind to CDK4 contained multiple amino acid changes, and these alterations were observed throughout the entire gene with no apparent mutational patterns or hot spots. Some of the mutations that moderately reduced the binding activity severely affected the kinase-inhibitory activity of p16.
Yang
R
R
Division of Hematology/Oncology, Cedars-Sinai Research Institute, UCLA School of Medicine 90048, USA.
Serrano
M
M
Slater
J
J
Leung
E
E
Koeffler
H P
HP
D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D017124:Conserved Sequence; D051358:Cyclin-Dependent Kinase 4; D019941:Cyclin-Dependent Kinase Inhibitor p16; D018844:Cyclin-Dependent Kinases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism*; D005800:Genes, Fungal; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017354:Point Mutation; D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
D002352:Carrier Proteins; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004791:Enzyme Inhibitors; D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases