Molecular cloning and characterization of the human p19INK4d gene promoter

FEBS Letters

Miyazawa, Kazuhiro²  Hitomi, Toshiaki²  Sakai, Toshiyuki²  Matsuzaki, Youichirou²  Yokota, Tomoya²  Yamagishi, Hisakazu¹ 

[1] Department of Digestive Surgery, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan;Department of Preventive Medicine, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan

关键词: p19INK4d;    Promoter;    Luciferase assay;    AP-2;    activating protein 2;    SRY;    sex determining region Y;   

DOI  :  10.1016/S0014-5793(02)02647-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

p19^INK4d, a member of the INK4 family of cyclin-dependent kinase (CDK) inhibitors, negatively regulates the cyclin D–CDK4/6 complexes, which promote G1/S transition by phosphorylating the retinoblastoma tumor-suppressor gene product. To investigate the mechanism of transcriptional regulation of the p19^INK4d gene, we characterized the 5′-flanking region of the human p19^INK4d gene. The cap-site hunting method revealed that the transcription starts at −16 nucleotide (nt) upstream of the initiation codon. The 5′-flanking region of the human p19^INK4d gene was ligated to a luciferase reporter gene and possessed functional promoter activity. Luciferase assay with a series of truncated 5′-flanking regions indicated that the region from −81 to −2 nt could drive the transcription of the p19^INK4d gene. Several Sp1 and activating protein 2 binding sites are located within the region from −81 to −2 nt. Mutation of the second Sp1 binding site from −33 to −25 nt decreased the promoter activity. Collectively, it was demonstrated that the human p19^INK4d gene is under the control of TATA-less promoter and the Sp1 binding site is involved in the transcription.

【 授权许可】

Unknown