Role of tropomyosin in formin-mediated contractile ring assembly in fission yeast.
Like animal cells, fission yeast divides by assembling actin filaments into a contractile ring. In addition to formin Cdc12p and profilin, the single tropomyosin isoform SpTm is required for contractile ring assembly. Cdc12p nucleates actin filaments and remains processively associated with the elongating barbed end while driving the addition of profilin-actin. SpTm is thought to stabilize mature filaments, but it is not known how SpTm localizes to the contractile ring and whether SpTm plays a direct role in Cdc12p-mediated actin polymerization. Using "bulk" and single actin filament assays, we discovered that Cdc12p can recruit SpTm to actin filaments and that SpTm has diverse effects on Cdc12p-mediated actin assembly. On its own, SpTm inhibits actin filament elongation and depolymerization. However, Cdc12p completely overcomes the combined inhibition of actin nucleation and barbed end elongation by profilin and SpTm. Furthermore, SpTm increases the length of Cdc12p-nucleated actin filaments by enhancing the elongation rate twofold and by allowing them to anneal end to end. In contrast, SpTm ultimately turns off Cdc12p-mediated elongation by "trapping" Cdc12p within annealed filaments or by dissociating Cdc12p from the barbed end. Therefore, SpTm makes multiple contributions to contractile ring assembly during and after actin polymerization.
Skau
Colleen T
CT
Department of Molecular Genetics, The University of Chicago, IL 60637, USA.
Neidt
Erin M
EM
Kovar
David R
DR
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D050296:Microbial Viability; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D011485:Protein Binding; D012568:Schizosaccharomyces / Q000166:cytology / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D014335:Tropomyosin / Q000378:metabolism*
D000199:Actins; C531994:Cdc12 protein, S pombe; C410737:Cdc8 protein, S pombe; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D029702:Schizosaccharomyces pombe Proteins; D014335:Tropomyosin; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate