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Arch Virol ; 147(1): 103-17, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11855624

RESUMO

The movement proteins BC1 and BV1 of Abutilon mosaic geminivirus fused to glutathion-S-transferase (GST) and Flag-peptide were expressed in fission yeast (Schizosaccharomyces pombe) cells to analyse the fundamental intracellular distribution of these proteins in an eukaryotic cell in the absence of plant-specific factors. Most of BC1 protein sedimented rapidly after cell lysis and differential centrifugation. Using freeze-fracture immuno-labelling, the protein was detected in situ predominantly at plasma membranes and to a lower extent at cytoplasmic vesicles but not in the cytoplasm, the nuclei, or the mitochondria. Anti-BC1, anti-GST, and anti-Flag antibodies tagged smooth flecks only at the protoplasmic faces of the plasma membrane. The consequences of the BC1 behaviour for its use in two-hybrid analysis in yeast are discussed. In contrast, BV1 was detected mainly in the nucleus and partially in the cytoplasm but never associated with membranes.


Assuntos
Membrana Celular/química , Vesículas Citoplasmáticas/química , Técnica de Fratura por Congelamento , Geminiviridae , Schizosaccharomyces/química , Frações Subcelulares/química , Proteínas Virais/análise , Membrana Celular/ultraestrutura , Clonagem Molecular , Vesículas Citoplasmáticas/ultraestrutura , Geminiviridae/genética , Glutationa Transferase , Microscopia Eletrônica , Proteínas do Movimento Viral em Plantas , Proteínas Recombinantes de Fusão/química , Schizosaccharomyces/genética , Schizosaccharomyces/ultraestrutura , Frações Subcelulares/ultraestrutura , Proteínas Virais/genética
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