Abstract
Fusion proteins between the green fluorescent protein (GFP) and the cytoskeleton proteins Act1p (actin), Sac6p (yeast fimbrin homolog), and Abp1p in budding yeast (Saccharomyces cerevisiae) localize to the cortical actin patches. The actin fusions could not function as the sole actin source in yeast, but fusions between the actin-binding proteins Abp1p and Sac6p complement fully the phenotypes associated with their gene deletions. Direct observation in vivo reveals that the actin cortical patches move. Movement of actin patches is constrained to the asymmetric distribution of the patches in growing cells, and this movement is greatly reduced when metabolic inhibitors such as sodium azide are added. Fusion protein-labeled patches are normally distributed during the yeast cell cycle and during mating. In vivo observation made possible the visualization of actin patches during sporulation as well.
MeSH Terms
Actins/analysis,biosynthesis,metabolism
Cytoskeleton/physiology,ultrastructure
Gene Deletion
Genes, Fungal
Genetic Complementation Test
Genotype
Green Fluorescent Proteins
Luminescent Proteins/analysis,biosynthesis
Microfilament Proteins/biosynthesis,metabolism
Models, Biological
Plasmids
Recombinant Fusion Proteins/analysis,biosynthesis,metabolism
Saccharomyces cerevisiae/cytology,genetics,physiology
Chemicals
Actins
Luminescent Proteins
Microfilament Proteins
Recombinant Fusion Proteins
Green Fluorescent Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Doyle T
Department of Genetics, Stanford University Medical Center, CA 94305-5120, USA.
Botstein D
References (20)
20 references, click to expand
-
Isolation and sequence of the gene for actin in Saccharomyces cerevisiae.
Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912-6
PMID: 7001447
-
GFP in mammalian cells.
Trends Genet. 1995 Aug;11(8):326-7
PMID: 8585132
-
Phenotypic analysis of temperature-sensitive yeast actin mutants.
Cell. 1985 Feb;40(2):405-16
PMID: 3967297
-
Plasmid construction by homologous recombination in yeast.
Gene. 1987;58(2-3):201-16
PMID: 2828185
-
Tubulin and actin topology during zygote formation of Saccharomyces cerevisiae.
J Gen Microbiol. 1987 Dec;133(12):3355-63
PMID: 3332685
-
Yeast actin-binding proteins: evidence for a role in morphogenesis.
J Cell Biol. 1988 Dec;107(6 Pt 2):2551-61
PMID: 3060468
-
A yeast actin-binding protein is encoded by SAC6, a gene found by suppression of an actin mutation.
Science. 1989 Jan 13;243(4888):231-3
PMID: 2643162
-
The SPA2 gene of Saccharomyces cerevisiae is important for pheromone-induced morphogenesis and efficient mating.
J Cell Biol. 1990 Oct;111(4):1451-64
PMID: 2211820
-
Primary structure of the Aequorea victoria green-fluorescent protein.
Gene. 1992 Feb 15;111(2):229-33
PMID: 1347277
-
Actin- and tubulin-dependent functions during Saccharomyces cerevisiae mating projection formation.
Mol Biol Cell. 1992 Apr;3(4):429-44
PMID: 1498363
-
Systematic mutational analysis of the yeast ACT1 gene.
Genetics. 1992 Oct;132(2):337-50
PMID: 1427032
-
end3 and end4: two mutants defective in receptor-mediated and fluid-phase endocytosis in Saccharomyces cerevisiae.
J Cell Biol. 1993 Jan;120(1):55-65
PMID: 8380177
-
Synthetic-lethal interactions identify two novel genes, SLA1 and SLA2, that control membrane cytoskeleton assembly in Saccharomyces cerevisiae.
J Cell Biol. 1993 Aug;122(3):635-44
PMID: 8335689
-
Green fluorescent protein as a marker for gene expression.
Science. 1994 Feb 11;263(5148):802-5
PMID: 8303295
-
Ultrastructure of the yeast actin cytoskeleton and its association with the plasma membrane.
J Cell Biol. 1994 Apr;125(2):381-91
PMID: 8163554
-
The yeast actin cytoskeleton.
Curr Opin Cell Biol. 1994 Feb;6(1):110-9
PMID: 8167016
-
Mapping actin surfaces required for functional interactions in vivo.
J Cell Biol. 1994 Jul;126(2):423-32
PMID: 8034743
-
Improved green fluorescence.
Nature. 1995 Feb 23;373(6516):663-4
PMID: 7854443
-
Understanding, improving and using green fluorescent proteins.
Trends Biochem Sci. 1995 Nov;20(11):448-55
PMID: 8578587
-
Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae.
J Cell Biol. 1984 Mar;98(3):934-45
PMID: 6365931