Detection of spectrin-like protein in yeasts: spectrin-actin
relations

D 2002

Detection of spectrin-like protein in yeasts: spectrin-actin relations

SLANINOVÁ, Iva, Alena HOLUBÁŘOVÁ and Augustin SVOBODA

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

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14110/02:00006732

Organization unit

Faculty of Medicine

Keywords in English

yeast; spectrin; cytoskeleton; actin

Abstract

V originále

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 project

Name: 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 project

Name: Úloha cytoskeletu a buněčné stěny v polarizaci buňky

Investor: Czech Science Foundation, The role of cytoskeleton and cell wall in the cell polarization

Citovat

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, 2002, p. 14.

@inproceedings{406732,
   author = {Slaninová, Iva and Holubářová, Alena and Svoboda, Augustin},
   address = {Praha},
   booktitle = {Sborník X. Cytoskeletálního klubu},
   keywords = {yeast; spectrin; cytoskeleton; actin},
   language = {eng},
   location = {Praha},
   pages = {14-14},
   publisher = {Československá biologická společnost},
   title = {Detection of spectrin-like protein in yeasts: spectrin-actin relations},
   year = {2002}
}

TY  - JOUR
ID  - 406732
AU  - Slaninová, Iva - Holubářová, Alena - Svoboda, Augustin
PY  - 2002
TI  - Detection of spectrin-like protein in yeasts: spectrin-actin relations
PB  - Československá biologická společnost
CY  - Praha
KW  - yeast
KW  - spectrin
KW  - cytoskeleton
KW  - actin
N2  - 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.
ER  -

SLANINOVÁ, Iva, Alena HOLUBÁŘOVÁ and Augustin SVOBODA. Detection of spectrin-like protein in yeasts: spectrin-actin relations. In \textit{Sborník X. Cytoskeletálního klubu}. Praha: Československá biologická společnost, 2002, p.~14.

Detailed Information on Publication Record