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J. Biol. Chem., Vol. 275, Issue 22, 16752-16757, June 2, 2000
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From the BSAP (Pax5) is an essential
transcription factor for early B cell and central nervous system
development. In later B cell development, BSAP has been implicated in
the regulation of 3' Ig enhancers and a number of B cell-specific
genes. Previous studies have suggested a role for BSAP-interacting
proteins in the regulation of the function of BSAP. Using the yeast
two-hybrid system, we identified importin
BSAP (Pax5)-Importin
1 (Rch1) Interaction
Identifies a Nuclear Localization Sequence*
§,,¶,, and
Department of Cell Biology, Albert Einstein
College of Medicine, Bronx, New York 10461 and the
§ Biology Department, Long Island University,
Brooklyn, New York 11201
1 (Rch1) as a
BSAP-interacting protein. Importin proteins have been shown to
escort proteins into the nucleus through interaction with a nuclear
localization signal (NLS), composed of short stretches of basic amino
acids. A predicted NLS in BSAP (NKRKRDE, located at amino acids
195-201 in the central domain) was confirmed to be essential for
interaction with importin 1 by the yeast two-hybrid assay. Physical
interaction between BSAP and importin 1 was detected in
vitro by a glutathione S-transferase (GST) pulldown
assay. The NLS sequence in BSAP conferred nuclear localization to green
fluorescent protein (GFP)-BSAP fusion proteins. Although the N-terminal
paired (DNA-binding) domain of BSAP also conferred nuclear localization
when coupled to green fluorescent protein, this domain did not bind to
importin 1 in the yeast two-hybrid assay. The NLS sequence in the
central domain of BSAP binds to the C-terminal 98-amino acid fragment
of importin 1.
*
This work was supported by National Institutes of Health
Grant R37AI13509 (to B. K. B.), Albert Einstein College of Medicine Cancer Center Grant PC30CA13330, and an American Society of Cell Biology-Minorities Affairs Committee Visiting Professorship Award (to
C. R. K.).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. Tel.:
718-430-2291; Fax: 718-430-8574; E-mail: birshtei@aecom.yu.edu.
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