title: Chimeras of p14ARF and p16: functional hybrids with the ability to arrest growth.
abstract: The INK4A locus codes for two independent tumor suppressors, p14ARF and p16/CDKN2A, and is frequently mutated in many cancers. Here we report a novel deletion/substitution from CC to T in the shared exon 2 of p14ARF/p16 in a melanoma cell line. This mutation aligns the reading frames of p14ARF and p16 mid-transcript, producing one protein which is half p14ARF and half p16, chimera ARF (chARF), and another which is half p16 and half non-p14ARF/non-p16 amino acids, p16-Alternate Carboxyl Terminal (p16-ACT). In an effort to understand the cellular impact of this novel mutation and others like it, we expressed the two protein products in a tumor cell line and analyzed common p14ARF and p16 pathways, including the p53/p21 and CDK4/cyclin D1 pathways, as well as the influence of the two proteins on growth and the cell cycle. We report that chARF mimicked wild-type p14ARF by inducing the p53/p21 pathway, inhibiting cell growth through G2/M arrest and maintaining a certain percentage of cells in G1 during nocodazole-induced G2 arrest. chARF also demonstrated p16 activity by binding CDK4. However, rather than preventing cyclin D1 from binding CDK4, chARF stabilized this interaction through p21 which bound CDK4. p16-ACT had no p16-related function as it was unable to inhibit cyclin D1/CDK4 complex formation and was unable to arrest the cell cycle, though it did inhibit colony formation. We conclude that these novel chimeric proteins, which are very similar to predicted p16/p14ARF chimeric proteins found in other primary cancers, result in maintained p14ARF-p53-p21 signaling while p16-dependent CDK4 inhibition is lost.
authors: Department of Pediatric Hematology/Oncology, University of California San Diego, San Diego, California, United States of America.
authors: Richard T
authors: RT
authors: Williams
authors: Department of Pediatric Hematology/Oncology, University of California San Diego, San Diego, California, United States of America.
authors: Lisa M
authors: LM
authors: Barnhill
authors: Genomics Research Center, Academia Sinica, Taipei, Taiwan.
authors: Huan-Hsien
authors: HH
authors: Kuo
authors: Genomics Research Center, Academia Sinica, Taipei, Taiwan.
authors: Wen-Der
authors: WD
authors: Lin
authors: Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, California, United States of America.
authors: Ayse
authors: A
authors: Batova
authors: Department of Pediatric Hematology/Oncology, University of California San Diego, San Diego, California, United States of America.
authors: Alice L
authors: AL
authors: Yu
authors: Department of Pediatric Hematology/Oncology, University of California San Diego, San Diego, California, United States of America.
authors: Mitchell B
authors: MB
authors: Diccianni
mesh_terms: D001483:Base Sequence; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019938:Cyclin D1 / Q000378:metabolism; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D006801:Humans; D008545:Melanoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D050506:Mutant Chimeric Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D025803:Tumor Suppressor Protein p14ARF / Q000235:genetics* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D050506:Mutant Chimeric Proteins; D025803:Tumor Suppressor Protein p14ARF; D016159:Tumor Suppressor Protein p53; D019938:Cyclin D1; D051358:Cyclin-Dependent Kinase 4