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Molecular and Cellular Biology, July 2000, p. 5140-5148, Vol. 20, No. 14
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Evidence for Gal3p's Cytoplasmic Location and
Gal80p's Dual Cytoplasmic-Nuclear Location Implicates New
Mechanisms for Controlling Gal4p Activity in
Saccharomyces cerevisiae
Gang
Peng1 and
James E.
Hopper1,2,*
Department of Biochemistry and Molecular
Biology1 and Intercollege Graduate
Program in Genetics,2 College of Medicine,
The Pennsylvania State University, Hershey, Pennsylvania 17033
Received 1 October 1999/Returned for modification 8 November
1999/Accepted 12 April 2000
Genetics and in vitro studies have shown that the direct
interaction between Gal3p and Gal80p plays a central role in
galactose-dependent Gal4p-mediated GAL gene expression in
the yeast Saccharomyces cerevisiae. Precisely how
Gal3p-Gal80p interaction effects induction is not clear. It has been
assumed that Gal3p interacts with Gal80p in the nucleus upon galactose
addition to release Gal80p inhibition of Gal4p. Although Gal80p has
been shown to possess nuclear localization signal (NLS) peptides, the
subcellular distribution of neither Gal80p nor Gal3p was previously
determined. Here we report that Gal3p is located in the cytoplasm and
apparently excluded from the nucleus. We show that Gal80p is located in
both the cytoplasm and the nucleus. Converting Gal80p into a
nucleus-localized protein (NLS-Gal80p) by exogenous NLS addition
impairs GAL gene induction. The impaired induction can be
partially suppressed by targeting Gal3p to the nucleus (NLS-Gal3p). We
document a very rapid association between NLS-Gal3p and Gal80p in vivo
in response to galactose, illustrating that the nuclear import of
Gal80p is very rapid and efficient. We also demonstrate that
nucleus-localized NLS-Gal80p can move out of the nucleus and shuttle
between nuclei in yeast heterokaryons. These results are the first
indication that the subcellular distribution dynamics of the Gal3 and
Gal80 proteins play a role in regulating Gal4p-mediated GAL
gene expression in vivo.
*
Corresponding author. Mailing address: Department of
Biochemistry and Molecular Biology, H171, The Pennsylvania State
University College of Medicine, 500 University Dr., Hershey, PA 17033. Phone: (717) 531-8590. Fax: (717) 531-7072. E-mail:
jhopper{at}psu.edu.
Molecular and Cellular Biology, July 2000, p. 5140-5148, Vol. 20, No. 14
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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