Zac1, a novel seven-zinc-finger transcription factor, preferentially binds GC-rich DNA elements and has intrinsic transactivation activity. To date, the NLS (nuclear localization signal) of Zac1 has not been empirically determined. We generated a series of EGFP (enhanced green fluorescence protein)-tagged deletion mutants of Zac1 and examined their subcellular localization, from which we defined two NLSs within the DNA-binding (or zinc-finger) domain. Fusion proteins consisting of the two EGFP-tagged zinc-finger clusters (zinc finger motifs 1–3 and 4–7) were located exclusively in the nucleus, demonstrating that each of the zinc-finger clusters is sufficient for nuclear localization. Physical interactions between these two zinc-finger clusters and importin α1 were demonstrated using an in vitro glutathione S-transferase pull-down assay. Finally, our results indicate that the association of Zac1 with importin α1 is also involved in regulating the transactivation activity of Zac1 on the p21WAF1/CIP1 gene and protein expression.
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Research Article|
February 12 2007
Importin α1 is involved in the nuclear localization of Zac1 and the induction of p21WAF1/CIP1 by Zac1
Shih-Ming Huang;
Shih-Ming Huang
1
1Department of Biochemistry and Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan 114, Republic of China
1To whom correspondence should be addressed (email shihming@ndmctsgh.edu.tw).
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Sheng-Ping Huang;
Sheng-Ping Huang
1Department of Biochemistry and Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan 114, Republic of China
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Sung-Ling Wang;
Sung-Ling Wang
1Department of Biochemistry and Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan 114, Republic of China
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Pei-Yao Liu
Pei-Yao Liu
1Department of Biochemistry and Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan 114, Republic of China
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Publisher: Portland Press Ltd
Received:
August 24 2006
Revision Received:
November 06 2006
Accepted:
November 16 2006
Accepted Manuscript online:
November 16 2006
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2007
Biochem J (2007) 402 (2): 359–366.
Article history
Received:
August 24 2006
Revision Received:
November 06 2006
Accepted:
November 16 2006
Accepted Manuscript online:
November 16 2006
Citation
Shih-Ming Huang, Sheng-Ping Huang, Sung-Ling Wang, Pei-Yao Liu; Importin α1 is involved in the nuclear localization of Zac1 and the induction of p21WAF1/CIP1 by Zac1. Biochem J 1 March 2007; 402 (2): 359–366. doi: https://doi.org/10.1042/BJ20061295
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