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J. Biol. Chem., Vol. 277, Issue 35, 32243-32252, August 30, 2002
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From the Department of Molecular, Cell and Developmental Biology,
Mount Sinai School of Medicine, New York, New York 10029
Erythroid Krüppel-like Factor (EKLF/KLF-1)
is an erythroid-specific transcription factor that contains three
C2H2 zinc fingers and is required for
correct chromatin structure and expression of the
Krüppel-like Zinc Fingers Bind to Nuclear Import Proteins
and Are Required for Efficient Nuclear Localization of Erythroid
Krüppel-like Factor*
-globin locus.
However, regions within the EKLF protein that serve as signals for its
nuclear localization and the proteins that may enable it to become
localized are unknown. Two approaches were used to address these
issues. First, green fluorescent protein or pyruvate kinase was fused
to EKLF domains, and localization was monitored and quantitated by
confocal microscopy. Two necessary and sufficient nuclear localization
signals (NLSs) were identified: one (NLS1) adjacent to the zinc finger
DNA binding domain within a highly basic stretch of amino acids
(275-296), and another more efficient signal (NLS2) within the zinc
finger domain itself (amino acids 293-376). Interestingly, each zinc
finger contributes to the overall effectiveness of NLS2 and requires an
intact finger structure. Second, each NLS was tested in
vitro for binding to importin proteins. Surprisingly, both EKLF
NLSs, but principally the zinc finger domain, bind importin 
and
importin . These findings demonstrate that two nuclear localization
signals target EKLF to the nucleus and suggest this transport relies
primarily on a novel zinc finger/importin protein interaction.
*
This work was supported by National Institutes of Health
Shared Instrumentation Grant 1 S10 RR0 9145-01 and National Science Foundation Major Research Instrumentation Grant DBI-9724504 and by
United States Public Health Services Grants T32 GM08553 (to K. J. Q.)
and R01 DK46865 (to J. J. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Molecular,
Cell and Developmental Biology, Mount Sinai School of Medicine, Box
1020, One Gustave L. Levy Place, New York, NY 10029. Tel.: 212-241-4143; Fax: 212-860-9279; E-mail: james.bieker@mssm.edu.
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