SLANINOVÁ, Iva, Alena HOLUBÁŘOVÁ and Augustin SVOBODA. Detection of spectrin-like protein in yeasts: spectrin-actin relations. In Sborník X. Cytoskeletálního klubu. Praha: Československá biologická společnost. p. 14. 2002.
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Basic information
Original name Detection of spectrin-like protein in yeasts: spectrin-actin relations
Authors SLANINOVÁ, Iva (203 Czech Republic, guarantor), Alena HOLUBÁŘOVÁ (203 Czech Republic) and Augustin SVOBODA (203 Czech Republic).
Edition Praha, Sborník X. Cytoskeletálního klubu, p. 14-14, 2002.
Publisher Československá biologická společnost
Other information
Original language English
Type of outcome Proceedings paper
Field of Study 10600 1.6 Biological sciences
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
RIV identification code RIV/00216224:14110/02:00006732
Organization unit Faculty of Medicine
Keywords in English yeast; spectrin; cytoskeleton; actin
Tags Actin, Cytoskeleton, spectrin, Yeast
Changed by Changed by: prof. MUDr. Iva Slaninová, Ph.D., učo 2105. Changed: 6/5/2003 16:22.
Abstract
Proteins related to the a- and b- spectrin have been detected in almost all vertebrate tissues, and also in plants (1) and hyphal fungae (2). We studied spectrin-like proteins in budding yeasts Saccharomyces cerevisiae and fission yeasts Schizosaccharomyces japonicus var. versatilis. Western blot analysis using anti-chicken and anti-human erythrocyte spectrin antibodies detected band at 220 kDa in cells and protoplasts extracts of both yeast strains. Additional bands were present at 50, 34 and 20 kDa in S. cerevisiae, and 44, 34 and 20 kDa in Sch. versatilis. Fluorescence microscopy revealed distinct labelling of the spectrin-like antigens at the cell surfaces and in vacuolar membranes of cells and protoplasts of both yeast species. In addition, the antigens were spread in the cytoplasm of both cells as fluorescent patches and as a diffuse fluorescence in Sch. versatilis nuclei. In mating and sporulating cells spectrin-like antigens lined the cell surface and prespore membranes. Electron microscopy of immunogold labelled cells and protoplasts revealed spectrin-like antigens in the cell wall, plasma membrane, ER, vacuole, nuclei, vesicles and mitochondria. Contrary to plant and some other fungal cells, spectrin in yeasts did not co-localize with actin. Growing regions of the cells were not preferentially labelled. Spectrin antigens correlated with actin only in protoplasts and maturing spores where patches of both lined the whole surface. The topology of spectrin was not affected by actin depolymerisation with Latrunculin B, nor was it changed in either act1-1 or cdc42 mutants under restrictive conditions. Under osmotic stress, both spectrin and actin were delocalized and appeared in the form of large clusters in the cytoplasm. Our results confirmed the role for spectrin as a membrane protein and the fact that spectrin localization occurs independently of actin distribution.
Links
GA204/00/0394, research and development projectName: Buněčná stěna kvasinek jako extracelulární matrix: cytoskelet a stěnové funkce
Investor: Czech Science Foundation, The yeast cell wall as an extracellular matrix: cytoskeleton and cell wall functions
GA204/99/D025, research and development projectName: Úloha cytoskeletu a buněčné stěny v polarizaci buňky
Investor: Czech Science Foundation, The role of cytoskeleton and cell wall in the cell polarization
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