Profilin binding to poly-L-proline and actin monomers along with ability to catalyze actin nucleotide exchange is required for viability of fission yeast.
We tested the ability of 87 profilin point mutations to complement temperature-sensitive and null mutations of the single profilin gene of the fission yeast Schizosaccharomyces pombe. We compared the biochemical properties of 13 stable noncomplementing profilins with an equal number of complementing profilin mutants. A large quantitative database revealed the following: 1) in a profilin null background fission yeast grow normally with profilin mutations having >10% of wild-type affinity for actin or poly-L-proline, but lower affinity for either ligand is incompatible with life; 2) in the cdc3-124 profilin ts background, fission yeast function with profilin having only 2-5% wild-type affinity for actin or poly-L-proline; and 3) special mutations show that the ability of profilin to catalyze nucleotide exchange by actin is an essential function. Thus, poly-L-proline binding, actin binding, and actin nucleotide exchange are each independent requirements for profilin function in fission yeast.
Lu
J
J
Graduate Program in Cellular and Molecular Medicine, Johns Hopkins Medical School, Baltimore, Maryland 21205, USA.
Pollard
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TD
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002384:Catalysis; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000378:metabolism*; D010641:Phenotype; D051304:Profilins; D011817:Rabbits; D012568:Schizosaccharomyces / Q000254:growth & development*; D029702:Schizosaccharomyces pombe Proteins; D013696:Temperature
D000199:Actins; D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D008024:Ligands; D010455:Peptides; D051304:Profilins; D029702:Schizosaccharomyces pombe Proteins; C490133:cdc3 protein, S pombe; C011083:polyproline