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PMID: 8756649 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The EH-domain-containing protein Pan1 is required for normal organization of the actin cytoskeleton in Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 16 ·No. 9 ·1996-09-00 ·Pages 4897-914

Tang HY, Cai M

Abstract

Normal cell growth and division in the yeast Saccharomyces cerevisiae involve dramatic and frequent changes in the organization of the actin cytoskeleton. Previous studies have suggested that the reorganization of the actin cytoskeleton in accordance with cell cycle progression is controlled, directly or indirectly, by the cyclin-dependent kinase Cdc28. Here we report that by isolating rapid-death mutants in the background of the Start-deficient cdc28-4 mutation, the essential yeast gene PAN1, previously thought to encode the yeast poly(A) nuclease, is identified as a new factor required for normal organization of the actin cytoskeleton. We show that at restrictive temperature, the pan1 mutant exhibited abnormal bud growth, failed to maintain a proper distribution of the actin cytoskeleton, was unable to reorganize actin the cytoskeleton during cell cycle, and was defective in cytokinesis. The mutant also displayed a random pattern of budding even at permissive temperature. Ectopic expression of PAN1 by the GAL promoter caused abnormal distribution of the actin cytoskeleton when a single-copy vector was used. Immunofluorescence staining revealed that the Pan1 protein colocalized with the cortical actin patches, suggesting that it may be a filamentous actin-binding protein. The Pan1 protein contains an EF-hand calcium-binding domain, a putative Src homology 3 (SH3)-binding domain, a region similar to the actin cytoskeleton assembly control protein Sla1, and two repeats of a newly identified protein motif known as the EH domain. These findings suggest that Pan1, recently recognized as not responsible for the poly(A) nuclease activity (A. B. Sachs and J. A. Deardorff, erratum, Cell 83:1059, 1995; R. Boeck, S. Tarun, Jr., M. Rieger, J. A. Deardorff, S. Muller-Auer, and A. B. Sachs, J. Biol. Chem. 271:432-438, 1996), plays an important role in the organization of the actin cytoskeleton in S. cerevisiae.

MeSH Terms
Actins/metabolism Amino Acid Sequence Cell Cycle Chitin/metabolism Cytoskeleton/ultrastructure Fungal Proteins/chemistry,genetics,physiology Microfilament Proteins Molecular Sequence Data Saccharomyces cerevisiae/metabolism,ultrastructure Saccharomyces cerevisiae Proteins Sequence Alignment Sequence Homology, Amino Acid src Homology Domains
Chemicals
Actins Fungal Proteins Microfilament Proteins PAN1 protein, S cerevisiae Saccharomyces cerevisiae Proteins Chitin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tang H Y
Institute of Molecular and Cell Biology, National University of Singapore.
Cai M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.9.4897
Published
1996-09-00
Pages
4897-914
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231492
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.16.9.4897 PMC Full Text View journal details
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