Electrophoretic analysis of the ribonucleoproteins of hepatitis delta virus
| Autor: | V V Bichko, Gloria Moraleda, Kate E. Dingle, John M. Taylor |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
viruses
Immunoblotting Biology Genome Virus Cell Line chemistry.chemical_compound Virology Animals Humans Ribonucleoprotein Electrophoresis Agar Gel Hepatitis delta Antigens Gel electrophoresis Aurintricarboxylic Acid Hepatitis Antigens virus diseases RNA Transfection biochemical phenomena metabolism and nutrition Blotting Northern Ribonucleoside Molecular biology Ribonucleoproteins chemistry Marmota Agarose Ribonucleosides Hepatitis Delta Virus Protein Binding |
| Zdroj: | Journal of Virological Methods. 75:199-204 |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/s0166-0934(98)00117-7 |
| Popis: | Replication of hepatitis delta virus (HDV) is dependent on delta antigen (deltaAg), an HDV-encoded protein, which binds to HDV RNA and is capable of multimerization. To characterize HDV-specific ribonucleoprotein complexes (RNP) we used electrophoresis into non-denaturing agarose gels followed by northern analysis, to detect HDV RNA, and immunoblot, to detect deltaAg. We studied RNP from three sources: (i) vRNP, disrupted virions obtained from infected woodchuck serum; (ii) sRNP, disrupted particles secreted from transfected cultured cells; and (iii) cRNP, isolated from cells in which HDV genome replication was occurring. sRNP were approximately 28% smaller than vRNP. Treatment of vRNP with aurin tricarboxylic acid disrupted both deltaAg-deltaAg and deltaAg-RNA interactions while vanadyl ribonucleosides released the RNA without causing detectable disruption of the multimeric deltaAg complex. cRNP were smaller and more heterogeneous than vRNP and sRNP, and probably contained host components. The application of these electrophoretic procedures, and especially the use of prior treatments with vanadyl ribonucleoside complexes have provided valuable information on the RNP of HDV, and we expect they should find applicability in RNP studies of other RNA viruses. |
| Databáze: | OpenAIRE |
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