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Originally published In Press as doi:10.1074/jbc.M208993200 on November 8, 2002
J. Biol. Chem., Vol. 278, Issue 2, 991-997, January 10, 2003
Nak1, an Essential Germinal Center (GC) Kinase Regulates Cell
Morphology and Growth in Schizosaccharomyces pombe*
Timothy Y.
Huang ,
Nancy A.
Markley§, and
Dallan
Young¶
From the Department of Biochemistry & Molecular Biology, University
of Calgary, Calgary, Alberta T2N 4N1, Canada
We have identified and characterized Nak1, a
652- amino acid NH2-terminal kinase belonging to
the group II germinal center kinase (GCK) family, in
Schizosaccharomyces pombe. We found that nak1
is essential for cell proliferation. Furthermore, partial repression of
nak1, under regulation of an integrated nmt1
promoter, resulted in an aberrant round cellular morphology, actin and
microtubule mislocalization, slow growth, and cell division defects.
Overexpression of either a kinase-inactive mutant
(Nak1K39R) or the non-catalytic domain resulted in similar
phenotypes, suggesting dominant-negative effects. By deletion analysis,
we mapped the region responsible for this dominant-negative effect to
the COOH-terminal 99 residues. Furthermore, we found that deletion of
the COOH-terminal 99 residues inhibited Nak1 autophosphorylation, and
expression of a partially inactive (Nak1T171A) or truncated
(Nak11-562) protein only weakly suppressed morphological
and growth phenotypes, indicating that both kinase and COOH-terminal
regions are important for Nak1 function. GFP-Nak1 localized uniformly
throughout the cytoplasm, unlike many other proteins which influence
cell polarity that preferentially localize to cell ends. Together, our
results implicate Nak1 in the regulation of cell polarity, growth, and division and suggest that the COOH-terminal end plays an important role
in the regulation of this kinase.
*
This work was supported by a grant (to D. Y.) from the
Canadian Institutes of Health Research.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.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF091345.
Supported by the National Sciences and Engineering Research
Council of Canada and the Alberta Heritage Foundation for Medical Research.
§
Supported by the Alberta Cancer Board.
¶
Supported by the Alberta Heritage Foundation for Medical
Research. To whom correspondence should be addressed: Dept. of
Biochemistry & Molecular Biology, University of Calgary, 3330 Hospital
Dr. NW, Calgary, Alberta T2N 4N1, Canada. Tel.: 403-220-3030; Fax: 403-283-8727; E-mail: young@ucalgary.ca.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

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