Immunodetection of Spectrin-like Proteins in Yeasts

J 2003

Immunodetection of Spectrin-like Proteins in Yeasts

SLANINOVÁ, Iva, Alena HOLUBÁŘOVÁ a Augustin SVOBODA

Základní údaje

Originální název

Immunodetection of Spectrin-like Proteins in Yeasts

Autoři

SLANINOVÁ, Iva (203 Česká republika, garant), Alena HOLUBÁŘOVÁ (203 Česká republika) a Augustin SVOBODA (203 Česká republika)

Vydání

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

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Kanada

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 1.094

Kód RIV

RIV/00216224:14110/03:00008399

Organizační jednotka

Lékařská fakulta

UT WoS

000183010400005

Klíčová slova anglicky

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

Štítky

Cytoskeleton, ELM, membrane skeleton, Saccharomyces, spectrin, Yeast

Změněno: 22. 6. 2009 12:10, prof. MUDr. Iva Slaninová, Ph.D.

Anotace

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.

Návaznosti

GA204/00/0394, projekt VaV

Název: Buněčná stěna kvasinek jako extracelulární matrix: cytoskelet a stěnové funkce

Investor: Grantová agentura ČR, Buněčná stěna kvasinek jako extracelulární matrix: cytoskelet a stěnové funkce

GA204/99/D025, projekt VaV

Název: Úloha cytoskeletu a buněčné stěny v polarizaci buňky

Investor: Grantová agentura ČR, Úloha cytoskeletu a buněčné stěny v polarizaci buňky

Citovat

SLANINOVÁ, Iva, Alena HOLUBÁŘOVÁ a 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, roč. 49, č. 3, s. 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 a 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, roč.~49, č.~3, s.~189-196. ISSN~0008-4166.

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