The integrity of the cytokinesis machinery under stress conditions requires the glucan synthase Bgs1p and its regulator Cfh3p.
In yeast, cytokinesis requires coordination between nuclear division, acto-myosin ring contraction, and septum synthesis. We studied the role of the Schizosaccharomyces pombe Bgs1p and Cfh3p proteins during cytokinesis under stress conditions. Cfh3p formed a ring in the septal area that contracted during mitosis; Cfh3p colocalized and co-immunoprecipitated with Cdc15p, showing that Cfh3p interacted with the contractile acto-myosin ring. In a wild-type strain, a significant number of contractile rings collapsed under stress conditions and this number increased dramatically in the cfh3Δ, bgs1cps1-191, and cfh3Δ bgs1/cps1-191. Our results show that after osmotic shock Cfh3p is essential for the stability of the (1,3) glucan synthase Bgs1p in the septal area, but not at the cell poles. Finally, cells adapted to stress; they repaired their contractile rings and re-localized Bgs1p to the cell surface some time after osmotic shock. A detailed analysis of the cytokinesis machinery in the presence of KCl revealed that the actomyosin ring collapsed before Bgs1p was internalized, and that it was repaired before Bgs1p re-localized to the cell surface. In the cfh3Δ, bgs1/cps1-191, and cfh3Δ bgs1/cps1-191 mutants, which have reduced glucan synthesis, the damage produced to the ring had stronger consequences, suggesting that an intact primary septum contributes to ring stability. The results show that the contractile actomyosin ring is very sensitive to stress, and that cells have efficient mechanisms to remedy the damage produced in this structure.
Departamento de Microbiología y Genética/Instituto de Biología Funcional y Genómica, Universidad de Salamanca/Consejo Superior de Investigaciones Científicas, Salamanca, Spain.
Mohammad Reza
MR
Sharifmoghadam
M-Ángeles
MÁ
Curto
Marta
M
Hoya
Nagore
N
de León
Rebeca
R
Martin-Garcia
Cristina
C
Doncel
M-Henar
MH
Valdivieso
D000199:Actins / Q000378:metabolism; D002455:Cell Division; D048749:Cytokinesis / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005964:Glucosyltransferases / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008957:Models, Genetic; D009154:Mutation; D009995:Osmosis; D010641:Phenotype; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013696:Temperature; D013997:Time Factors
D000199:Actins; C541035:Cfh3 protein, S pombe; D003598:Cytoskeletal Proteins; D029702:Schizosaccharomyces pombe Proteins; C513274:cdc4 protein, S pombe; D049452:Green Fluorescent Proteins; D005964:Glucosyltransferases; C020570:1,3-beta-glucan synthase