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J. Biol. Chem., Vol. 278, Issue 9, 6731-6740, February 28, 2003
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From the Department of Biochemistry, Mature poly(A) RNA transcripts are exported from
the nucleus in complex with heterogeneous nuclear ribonucleoproteins
(hnRNPs). Nab2p is an essential Saccharomyces cerevisiae
hnRNP protein that interacts with poly(A) RNA and shuttles between the
nucleus and cytoplasm. Functional Nab2p is required for export of
poly(A) RNA from the nucleus. The Nab2 protein consists of the
following four domains: a unique N-terminal domain, a glutamine-rich
domain, an arginine-glycine (RGG) domain, and a zinc finger domain. We generated Nab2p deletion mutants to analyze the contribution of each
domain to the in vivo function of Nab2p. We first tested whether the deletion mutants could replace the essential
NAB2 gene. We then examined the impact of these mutations
on Nab2p localization, poly(A) RNA localization, and association of
Nab2p with poly(A) RNA. Our analyses revealed that the N-terminal
domain is required for nuclear export of both poly(A) RNA and Nab2p. We
confirm that the RGG domain is important for Nab2p import in vivo. Finally, the zinc finger domain is critical for the
interaction between Nab2p and poly(A) RNA in vivo. Our data
support a model where Nab2p associates with poly(A) RNA in the nucleus
through the zinc finger domain and facilitates the export of the
poly(A) RNA through protein interactions mediated by the N-terminal domain.
Domain Analysis of the Saccharomyces cerevisiae
Heterogeneous Nuclear Ribonucleoprotein, Nab2p
DISSECTING THE REQUIREMENTS FOR Nab2p-FACILITATED POLY(A) RNA
EXPORT*
,
, and Graduate Program
in Genetics and Molecular Biology and ¶ Graduate Program in
Biochemistry, Cell, and Developmental Biology, Emory University School
of Medicine, Atlanta, Georgia 30322
*
This work was supported by a grant from the National
Institutes of Health (to A. H. C.).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.
Supported by the Emory Minority Graduate Fellowship.
**
To whom correspondence should be addressed: 4117 Rollins Research
Center, Emory University, 1510 Clifton Rd., N.E., Atlanta, GA
30322. Tel.: 404-727-4546; Fax: 404-727-3549; E-mail:
acorbe2@emory.edu.
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