Eukaryotic Cell
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Eukaryotic Cell, August 2008, p. 1246-1255, Vol. 7, No. 8
1535-9778/08/$08.00+0     doi:10.1128/EC.00024-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Saccharomyces cerevisiae Afr1 Protein Is a Protein Phosphatase 1/Glc7-Targeting Subunit That Regulates the Septin Cytoskeleton during Mating{triangledown}

Jennifer P. Bharucha,1,{dagger} Jennifer R. Larson,1 James B. Konopka,2 and Kelly Tatchell1*

Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130,1 Department of Molecular Genetics and Microbiology, State University of New York, Stony Brook, Stony Brook, New York 11794-52222

Received 18 January 2008/ Accepted 27 May 2008

Glc7, the type1 serine/threonine phosphatase in the yeast Saccharomyces cerevisiae, is targeted by auxiliary subunits to numerous locations in the cell, where it regulates a range of physiological pathways. We show here that the accumulation of Glc7 at mating projections requires Afr1, a protein required for the formation of normal projections. AFR1-null mutants fail to target Glc7 to projections, and an Afr1 variant specifically defective in binding to Glc7 [Afr1(V546A F548A)] forms aberrant projections. The septin filaments in mating projections of AFR1 mutants initiate normally but then rearrange asymmetrically as the projection develops, suggesting that the Afr1-Glc7 holoenzyme may regulate the maintenance of septin complexes during mating. These results demonstrate a previously unknown role for Afr1 in targeting Glc7 to mating projections and in regulating the septin architecture during mating.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, 1501 Kings Highway, LSU Health Sciences Center, Shreveport LA 71130. Phone: (318) 675-7769. Fax: (318) 675-5180. E-mail: ktatch{at}lsuhsc.edu

{triangledown} Published ahead of print on 13 June 2008.

{dagger} Present address: Laboratory of Human Retrovirology, Applied and Developmental Research Support Program, Science Application International Corporation (SAIC)—Frederick Inc., National Cancer Institute at Frederick, Frederick, MD 21702.

Eukaryotic Cell, August 2008, p. 1246-1255, Vol. 7, No. 8
1535-9778/08/$08.00+0     doi:10.1128/EC.00024-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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