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Titolo:
EFFECT OF BAFILOMYCIN A1 AND NOCODAZOLE ON ENDOCYTIC TRANSPORT IN HELA-CELLS - IMPLICATIONS FOR VIRAL UNCOATING AND INFECTION
Autore:
BAYER N; SCHOBER D; PRCHLA E; MURPHY RF; BLAAS D; FUCHS R;
Indirizzi:
UNIV VIENNA,DEPT GEN & EXPT PATHOL,WAEHRINGER GUERTEL 18-20 A-1090 VIENNA AUSTRIA UNIV VIENNA,DEPT GEN & EXPT PATHOL A-1090 VIENNA AUSTRIA UNIV VIENNA,INST BIOCHEM A-1090 VIENNA AUSTRIA CARNEGIE MELLON UNIV,DEPT BIOL SCI PITTSBURGH PA 15213
Titolo Testata:
Journal of virology (Print)
fascicolo: 12, volume: 72, anno: 1998,
pagine: 9645 - 9655
SICI:
0022-538X(1998)72:12<9645:EOBAAN>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
HUMAN RHINOVIRUS SEROTYPE-2; VACUOLAR H+-ATPASE; LATE ENDOSOMES; IN-VITRO; MULTIVESICULAR ENDOSOMES; VIRUS ENTRY; LOW-PH; ACIDIFICATION; LYSOSOMES; MATURATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
74
Recensione:
Indirizzi per estratti:
Citazione:
N. Bayer et al., "EFFECT OF BAFILOMYCIN A1 AND NOCODAZOLE ON ENDOCYTIC TRANSPORT IN HELA-CELLS - IMPLICATIONS FOR VIRAL UNCOATING AND INFECTION", Journal of virology (Print), 72(12), 1998, pp. 9645-9655

Abstract

Bafilomycin Al (baf), a specific inhibitor of vacuolar proton ATPases, is commonly employed to demonstrate the requirement of low endosomalpH for viral uncoating. However, in certain cell types baf also affects the transport of endocytosed material from early to late endocytic compartments. To characterize the endocytic route in HeLa cells that are frequently used to study early events in viral infection, we used S-35-labeled human rhinovirus serotype 2 (HRV2) together with various fluid-phase markers. These virions are taken up via receptor-mediated endocytosis and undergo a conformational change to C-antigenic particles at a pH of <5.6, resulting in release of the genomic RNA and ultimately in infection (E. Prchla, E. Kuechler, D. Blaas, and R. Fuchs, J. Virol. 68:3713-3723, 1994). As revealed by fluorescence microscopy and subcellular fractionation of microsomes by free-flow electrophoresis (FFE), baf arrests the transport of all markers in early endosomes. In contrast, the microtubule-disrupting agent nocodazole was found to inhibit transport by accumulating marker in endosomal carrier vesicles (ECV), a compartment intermediate between early and late endosomes. Accordingly, lysosomal degradation of HRV2 was suppressed, whereas its conformational change and infectivity remained unaffected by this drug. Analysis of the subcellular distribution of HRV2 and fluid-phase markers in the presence of nocodazole by FFE revealed no difference from thecontrol incubation in the absence of nocodazole. ECV and late endosomes thus have identical electrophoretic mobilities, and intraluminal pHs of <5.6 and allow uncoating of HRV2. As bafilomycin not only dissipates the low endosomal pH but also blocks transport from early to late endosomes in HeLa cells, its inhibitory effect on viral infection could in part also be attributed to trapping of virus in early endosomes which might lack components essential for uncoating. Consequently, inhibition of viral uncoating by bafilomycin cannot be taken to indicate alow pH requirement only.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 07/07/20 alle ore 22:07:07