A nucleus-localization-deficient mutant serves as a dominant-negative inhibitor of gut-enriched Krüppel-like factor function

Jue-Lon Shie, Chi-Chuan Tseng

Research output: Contribution to journalJournal Article peer-review

Abstract

Cancer cells differ from normal cells in many aspects, including loss of differentiation and uninhibited cell proliferation. Recent studies have suggested that gut-enriched Krüppel-like factor (GKLF) played an important role in the regulation of cell growth in the colon. Studies from this laboratory have shown that GKLF protein predominantly expressed in the cytoplasm but not the nucleus of colon cancer cells, suggesting that impaired nuclear translocation of GKLF might contribute to cancer formation. In this report, a region containing putative nuclear localization signal (NLS) of GKLF (PKRGRR; amino acids 385-390) was investigated. Mutation of KR to WT had no effect on the inhibitory properties of GKLF on cyclin D1 promoter activity and [3H]thymidine uptake in HT-29 cells, whereas mutation of RR to GL abolished GKLF function completely. Additional mutation analyses demonstrated that Arg390 is the most critical moiety within this region that mediated GKLF function and its nucleus localization. Cotransfection of Arg390 mutant (RR/RS) completely inhibited wild-type GKLF function, and GFP-RR/RS GKLF fusion proteins failed to translocate to the nucleus. The results from this study demonstrate that Arg390 confers the NLS of GKLF and that the nucleus-localization-deficient mutant serves as dominant-negative inhibitor of GKLF function. © 2001 Academic Press.
Original languageAmerican English
Pages (from-to)205-208
JournalBiochemical and Biophysical Research Communications
Volume283
Issue number1
DOIs
StatePublished - 2001

Keywords

  • Cyclin D1 promoter
  • GKLF
  • KLF4
  • NLS

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