Identification of the sequence responsible for the nuclear localization of human Cdc6

FEBS Letters

Yamamoto, Koshi²  Tsujimoto, Gozoh¹  Takei, Yoshinori¹ 

[1] Division of Molecular Cell Pharmacology, National Children's Medical Research Center, 3-35-31 Taishido, Setagaya-ku, Tokyo, 154-8509 Japan;Department of Pharmacology, Science University of Tokyo, 12 Funakawara-cho Ichigaya Sinzyuku-ku, Tokyo, 162-0826 Japan

关键词: Cdc6;    DNA replication;    Nuclear localization;    BSA;    bovine serum albumin;    DAB;    3;    3′-diaminobenzidine;    DTT;    dithiothreitol;    EGTA;    [ethylene bis(oxo-nitrilo)]tetraacetic acid;    GFP;    green fluorescent protein;    NLS;    nuclear localization signal;    GST;    glutathione S transferase;    nt;    nucleotide;    ORC;    origin recognition complex;    PAGE;    polyacrylamide gel electrophoresis;    SDS;    sodium dodecyl sulfate;    PBS;    phosphate-buffered saline;    PCR;    polymerase chain reaction;   

DOI  :  10.1016/S0014-5793(99)00306-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The Cdc6 is the essential protein for the initiation of DNA replication. Cdc6 is localized in the G1 nucleus, and abnormal nuclear localization of this protein induces irregular initiation of DNA replication. We identified here that amino acids K⁵⁷ and R⁵⁸ in the human Cdc6 protein play an important role in the nuclear localization of the protein. The fundamental features of the mechanism regulating the localization of Cdc6 seem to be maintained in yeast, Xenopus, and human, since the amino acid sequence surrounding K⁵⁷ and R⁵⁸, (S/T)PXKR(L/I), is conserved in these species. Substitution of amino acid residue S⁵⁴ with E and not Q blocked partially the nuclear localization of the protein, implying that the phosphorylation at S⁵⁴ is involved in the regulating mechanism of the cell cycle-dependent localization of Cdc6.

【 授权许可】

Unknown