An engineered site for protein kinase C flanking the SV40 large T‐antigen NLS confers phorbol ester‐inducible nuclear import

FEBS Letters

Jans, David A¹  Xiao, Chong-Yun¹ 

[1] Nuclear Signalling Laboratory, Division for Biochemistry and Molecular Biology, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia

关键词: Ca2+/phospholipid-dependent protein kinase C;    Nuclear import kinetics;    Confocal laser scanning microscopy;    Microinjection;    Phosphorylation-regulated nuclear localization sequence;   

DOI  :  10.1016/S0014-5793(98)01157-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Nuclear import of simian virus SV40 large tumour antigen (T-ag) is enhanced by the protein kinase CK2 (CK2) site flanking the nuclear localisation sequence (NLS). We report here that replacement of this site with a consensus site for protein kinase C (PK-C) can alter the regulation of T-ag nuclear import and render it inducible by phorbol ester. Measurement of nuclear import kinetics using fluorescently labelled proteins and confocal laser scanning microscopy show that the introduced PK-C site is functional in enhancing T-ag nuclear import compared to a protein lacking the CK2 site. Treatment with the PK-C activator phorbol 12-myristate 13-acetate (PMA) further increases the level of maximal nuclear accumulation and the initial nuclear import rate. This engineered PMA-responsive NLS may have application in targeting of molecules of interest to the nucleus in response to agents stimulating PK-C.

【 授权许可】

Unknown