Purification of profilin from Saccharomyces cerevisiae and analysis of profilin-deficient cells.
We have isolated profilin from yeast (Saccharomyces cerevisiae) and have microsequenced a portion of the protein to confirm its identity; the region microsequenced agrees with the predicted amino acid sequence from a profilin gene recently isolated from S. cerevisiae (Magdolen, V., U. Oechsner, G. Müller, and W. Bandlow. 1988. Mol. Cell. Biol. 8:5108-5115). Yeast profilin resembles profilins from other organisms in molecular mass and in the ability to bind to polyproline, retard the rate of actin polymerization, and inhibit hydrolysis of ATP by monomeric actin. Using strains that carry disruptions or deletions of the profilin gene, we have found that, under appropriate conditions, cells can survive without detectable profilin. Such cells grow slowly, are temperature sensitive, lose the normal ellipsoidal shape of yeast cells, often become multinucleate, and generally grow much larger than wild-type cells. In addition, these cells exhibit delocalized deposition of cell wall chitin and have dramatically altered actin distributions.
Haarer
B K
BK
Department of Anatomy and Cell Biology, University of Michigan Medical School, Ann Arbor 48109.
Lillie
S H
SH
Adams
A E
AE
Magdolen
V
V
Bandlow
W
W
Brown
S S
SS
D000199:Actins / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D003285:Contractile Proteins / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005800:Genes, Fungal; D007700:Kinetics; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D009154:Mutation; D051304:Profilins; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure
D000199:Actins; D003285:Contractile Proteins; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D051304:Profilins