The fission yeast Schizosaccharomyces pombe divides by medial fission and, like many higher eukaryotic cells, requires the function of an F-actin contractile ring for cytokinesis. In S. pombe, a class of cdc- mutants defective for cytokinesis, but not for DNA replication, mitosis, or septum synthesis, have been identified. In this paper, we present the characterization of one of these mutants, cdc3-124. Temperature shift experiments reveal that mutants in cdc3 are incapable of forming an F-actin contractile ring. We have molecularly cloned cdc3 and used the cdc3+ genomic DNA to create a strain carrying a cdc3 null mutation by homologous recombination in vivo. Cells bearing a cdc3-null allele are inviable. They arrest the cell cycle at cytokinesis without forming a contractile ring. DNA sequence analysis of the cdc3+ gene reveals that it encodes profilin, an actin-monomer-binding protein. In light of recent studies with profilins, we propose that Cdc3-profilin plays an essential role in cytokinesis by catalyzing the formation of the F-actin contractile ring. Consistent with this proposal are our observations that Cdc3-profilin localizes to the medial region of the cell where the F-actin contractile ring forms, and that it is essential for F-actin ring formation. Cells overproducing Cdc3-profilin become elongated, dumbbell shaped, and arrest at cytokinesis without any detectable F-actin staining. This effect of Cdc3-profilin overproduction is relieved by introduction of a multicopy plasmid carrying the actin encoding gene, act1+. We attribute these effects to potential sequestration of actin monomers by profilin, when present in excess.
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15 June 1994
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June 15 1994
The Schizosaccharomyces pombe cdc3+ gene encodes a profilin essential for cytokinesis.
M K Balasubramanian,
M K Balasubramanian
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
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B R Hirani,
B R Hirani
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
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J D Burke,
J D Burke
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
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K L Gould
K L Gould
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
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M K Balasubramanian
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
B R Hirani
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
J D Burke
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
K L Gould
Department of Cell Biology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232.
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1994) 125 (6): 1289–1301.
Citation
M K Balasubramanian, B R Hirani, J D Burke, K L Gould; The Schizosaccharomyces pombe cdc3+ gene encodes a profilin essential for cytokinesis.. J Cell Biol 15 June 1994; 125 (6): 1289–1301. doi: https://doi.org/10.1083/jcb.125.6.1289
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