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© The Rockefeller University Press, 0021-9525/1999//1203 $5.00
The Journal of Cell Biology, Volume 144, Number 6, , 1999 1203-1218

Novel Protein Kinases Ark1p and Prk1p Associate with and Regulate the Cortical Actin Cytoskeleton in Budding Yeast

Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3202

Ark1p (actin regulating kinase 1) was identified as a yeast protein that binds to Sla2p, an evolutionarily conserved cortical actin cytoskeleton protein. Ark1p and a second yeast protein, Prk1p, contain NH₂-terminal kinase domains that are 70% identical. Together with six other putative kinases from a number of organisms, these proteins define a new protein kinase family that we have named the Ark family.

Lack of both Ark1p and Prk1p resulted in the formation of large cytoplasmic actin clumps and severe defects in cell growth. These defects were rescued by wild-type, but not by kinase-dead versions of the proteins. Elevated levels of either Ark1p or Prk1p caused a number of actin and cell morphological defects that were not observed when the kinase-dead versions were overexpressed instead. Ark1p and Prk1p were shown to localize to actin cortical patches, making these two kinases the first signaling proteins demonstrated to be patch components. These results suggest that Ark1p and Prk1p may be downstream effectors of signaling pathways that control actin patch organization and function. Furthermore, results of double-mutant analyses suggest that Ark1p and Prk1p function in overlapping but distinct pathways that regulate the cortical actin cytoskeleton.

Key Words: protein kinases • Saccharomyces cerevisiae • cyclin-G–associated kinases (GAKs) • endocytosis • actin

Abbreviations used in this paper: DIC, differential interference-contrast (Nomarski); GAK, cyclin-G–associated kinase; GFP, green fluorescent protein; ORF, open reading frame; SM, synthetic medium.

Shirley Yang's present address is Baylor College of Medicine, Department of Microbiology and Immunology, Houston, TX 77030.

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