Immunodetection of Spectrin-like Proteins in Yeasts

J 2003

Immunodetection of Spectrin-like Proteins in Yeasts

SLANINOVÁ, Iva, Alena HOLUBÁŘOVÁ and Augustin SVOBODA

Basic information

Original name

Immunodetection of Spectrin-like Proteins in Yeasts

Authors

SLANINOVÁ, Iva (203 Czech Republic, guarantor), Alena HOLUBÁŘOVÁ (203 Czech Republic) and Augustin SVOBODA (203 Czech Republic)

Edition

Canadian Journal of Microbiology, MONTREAL RD, OTTAWA, ONTARIO K1A 0R6, CA, NATL RESEARCH COUNCIL CANADA, 2003, 0008-4166

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Canada

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 1.094

RIV identification code

RIV/00216224:14110/03:00008399

Organization unit

Faculty of Medicine

UT WoS

000183010400005

Keywords in English

spectrin; yeast; cytoskeleton; ELM; Saccharomyces; membrane skeleton

Abstract

V originále

Spectrin, a component of the membrane skeleton in erythrocytes and other animal cells, has also been identified in plant and fungal cells. However, its postulated role, i.e., the maintenance of shape and elasticity of the plasma membrane, is probably not exerted in walled cells. To study spectrin in these cells, we chose yeasts because of a high morphological variability of their life cycle. The localization of spectrin in the cells and protoplasts of Saccharomyces cerevisiae and Schizosaccharomyces japonicus var. versatilis was detected by Western blotting, indirect immunofluorescence and immunogold electron microscopy techniques with the use of anti-chicken and anti-human erythrocyte spectrin antibodies. A 220/240 kDa protein band and some bands of lower molecular weights were detected in cell and protoplast extracts of both yeast strains. Spectrin-like proteins were revealed by fluorescence microscopy at cell surfaces and in vacuolar membranes. Immunogold-labelling showed spectrin-like proteins in the plasma membrane, endoplasmic reticulum, vacuoles, nuclei, vesicles, mitochondria and cell walls. The topology of spectrin was not affected by actin depolymerization 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. It is concluded that a protein cross-reacting with spectrin antibodies is present in fission and budding yeasts. Generally, it is located in the proximity of the plasma membrane and other intracellular membranes, probably as a part of the membrane skeleton. No evidence of its relationship to either actin or growth zones of the cell can be provided.

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. Immunodetection of Spectrin-like Proteins in Yeasts. Canadian Journal of Microbiology. MONTREAL RD, OTTAWA, ONTARIO K1A 0R6, CA: NATL RESEARCH COUNCIL CANADA, 2003, vol. 49, No 3, p. 189-196. ISSN 0008-4166.

@article{491135,
   author = {Slaninová, Iva and Holubářová, Alena and Svoboda, Augustin},
   article_location = {MONTREAL RD, OTTAWA, ONTARIO K1A 0R6, CA},
   article_number = {3},
   keywords = {spectrin; yeast; cytoskeleton; ELM; Saccharomyces; membrane skeleton},
   language = {eng},
   issn = {0008-4166},
   journal = {Canadian Journal of Microbiology},
   title = {Immunodetection of Spectrin-like Proteins in Yeasts},
   volume = {49},
   year = {2003}
}

TY  - JOUR
ID  - 491135
AU  - Slaninová, Iva - Holubářová, Alena - Svoboda, Augustin
PY  - 2003
TI  - Immunodetection of Spectrin-like Proteins in Yeasts
JF  - Canadian Journal of Microbiology
VL  - 49
IS  - 3
SP  - 189-196
EP  - 189-196
PB  - NATL RESEARCH COUNCIL CANADA
SN  - 00084166
KW  - spectrin
KW  - yeast
KW  - cytoskeleton
KW  - ELM
KW  - Saccharomyces
KW  - membrane skeleton
N2  - Spectrin, a component of the membrane skeleton in erythrocytes and other animal cells, has also been identified in plant and fungal cells. However, its postulated role, i.e., the maintenance of shape and elasticity of the plasma membrane, is probably not exerted in walled cells. To study spectrin in these cells, we chose yeasts because of a high morphological variability of their life cycle. The localization of spectrin in the cells and protoplasts of Saccharomyces cerevisiae and Schizosaccharomyces japonicus var. versatilis was detected by Western blotting, indirect immunofluorescence and immunogold electron microscopy techniques with the use of anti-chicken and anti-human erythrocyte spectrin antibodies. A 220/240 kDa protein band and some bands of lower molecular weights were detected in cell and protoplast extracts of both yeast strains. Spectrin-like proteins were revealed by fluorescence microscopy at cell surfaces and in vacuolar membranes. Immunogold-labelling showed spectrin-like proteins in the plasma membrane, endoplasmic reticulum, vacuoles, nuclei, vesicles, mitochondria and cell walls. The topology of spectrin was not affected by actin depolymerization 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. It is concluded that a protein cross-reacting with spectrin antibodies is present in fission and budding yeasts. Generally, it is located in the proximity of the plasma membrane and other intracellular membranes, probably as a part of the membrane skeleton. No evidence of its relationship to either actin or growth zones of the cell can be provided.
ER  -

SLANINOVÁ, Iva, Alena HOLUBÁŘOVÁ and Augustin SVOBODA. Immunodetection of Spectrin-like Proteins in Yeasts. \textit{Canadian Journal of Microbiology}. MONTREAL RD, OTTAWA, ONTARIO K1A 0R6, CA: NATL RESEARCH COUNCIL CANADA, 2003, vol.~49, No~3, p.~189-196. ISSN~0008-4166.

Detailed Information on Publication Record