Absence of R24C mutation of the CDK4 gene in leukemias and solid tumors

• Published in: International journal of hematology Vol. 77; no. 3; pp. 259 - 262
• Main Authors: MORI, Naoki, RONG YANG, KAWAMATA, Norihiko, MILLER, Carl W, MIZOGUCHI, Hideaki, KOEFFLER, H. Phillip
• Format: Journal Article
• Language: English
• Published: Tokyo Springer 01-04-2003; Springer Nature B.V
• Subjects: Biological and medical sciences; Cell Line, Tumor; Cell physiology; Cell transformation and carcinogenesis. Action of oncogenes and antioncogenes; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase Inhibitor p16 - metabolism; Cyclin-Dependent Kinases - genetics; Cyclin-Dependent Kinases - metabolism; DNA Mutational Analysis; Fundamental and applied biological sciences. Psychology; Humans; Leukemia - genetics; Molecular and cellular biology; Mutagenesis; Neoplasms - genetics; Point Mutation; Protein Binding - genetics; Proto-Oncogene Proteins; Two-Hybrid System Techniques; Human; Enzyme; Transferases; Leukemia; Malignant hemopathy; Malignant tumor; In vitro; G1 Phase; Gene; Protein kinase; Cell cycle; Mutation; Cyclin dependent kinase 4
• ISSN: 0925-5710; 1865-3774
• DOI: 10.1007/bf02983783

A mutation of the p16(INK4a)-binding domain of the cyclin dependent kinase 4 (CDK4) gene, R24C, has been reported in some cases of melanoma. This mutation prevented binding of the CDK4 inhibitor p16(INK48) to CDK4. To determine the relevance of the mutation, we performed polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis in diverse types of human leukemias and solid tumors. No mobility shifts indicating sequence alterations were observed in 273 tumors and 49 cell lines from diverse kinds of tumors These results suggest that in contrast to melanoma, in many other types of human neoplasms the mutation of the CDK4 gene is very rare. To better understand these findings, we randomly mutagenized the CDK4 gene and used the yeast two-hybrid method to screen for CDK4 mutants that had lost the ability to bind to p16(INK4a). Sequence analysis and in vitro kinase assays showed that most of the mutations that disrupted interactions with p16(INK4) also knocked out the activity of CDK4. This result may explain the rareness of CDK4 mutations in human tumors.