Nak1, an essential germinal center (GC) kinase regulates cell morphology and growth in Schizosaccharomyces pombe.
We have identified and characterized Nak1, a 652- amino acid NH(2)-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 (Nak1(K39R)) 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 (Nak1(T171A)) or truncated (Nak1(1-562)) 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.
Huang
Timothy Y
TY
Department of Biochemistry & Molecular Biology, University of Calgary, Alberta T2N 4N1, Canada.
Markley
Nancy A
NA
Young
Dallan
D
D000595:Amino Acid Sequence; D016764:Cell Polarity; D003593:Cytoplasm / Q000737:chemistry; D000081410:Germinal Center Kinases; D008969:Molecular Sequence Data; D017346:Protein Serine-Threonine Kinases / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D012568:Schizosaccharomyces / Q000166:cytology* / Q000254:growth & development; D029702:Schizosaccharomyces pombe Proteins / Q000502:physiology*
D000081410:Germinal Center Kinases; D029702:Schizosaccharomyces pombe Proteins; D017346:Protein Serine-Threonine Kinases