The essential yeast Tcp1 protein affects actin and microtubules.
Previously, we showed that the yeast Saccharomyces cerevisiae cold-sensitive mutation tcp1-1 confers growth arrest concomitant with cytoskeletal disorganization and disruption of microtubule-mediated processes. We have identified two new recessive mutations, tcp1-2 and tcp1-3, that confer heat- and cold-sensitive growth. Cells carrying tcp1 alleles were analyzed after exposure to the appropriate restrictive temperatures by cell viability tests, differential contrast microscopy, fluorescent, and immunofluorescent microscopy of DNA, tubulin, and actin and by determining the DNA content per cell. All three mutations conferred unique phenotypes indicative of cytoskeletal dysfunction. A causal relationship between loss of Tcp1p function and the development of cytoskeletal abnormalities was established by double mutant analyses. Novel phenotypes indicative of allele-specific genetic interactions were observed when tcp1-1 was combined in the same strain with tub1-1, tub2-402, act1-1, and act1-4, but not with other tubulin or actin mutations or with mutations in other genes affecting the cytoskeleton. Also, overproduction of wild-type Tcp1p partially suppressed growth defects conferred by act1-1 and act1-4. Furthermore, Tcp1p was localized to the cytoplasm and the cell cortex. Based on our results, we propose that Tcp1p is required for normal development and function of actin and microtubules either through direct or indirect interaction with the major cytoskeletal components.
Ursic
D
D
Laboratories of Molecular Biology and Genetics, University of Wisconsin, Madison 53706.
Sedbrook
J C
JC
Himmel
K L
KL
Culbertson
M R
MR
D000199:Actins / Q000378:metabolism*; D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D002453:Cell Cycle / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins; D003593:Cytoplasm / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal; D005808:Genes, Recessive; D051379:Mice; D008870:Microtubules / Q000378:metabolism*; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D014404:Tubulin / Q000235:genetics / Q000378:metabolism
D000199:Actins; D005656:Fungal Proteins; D018832:Molecular Chaperones; C533315:Tcp1 protein, mouse; D014404:Tubulin; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins