The antigen of hepatitis delta virus: examination of in vitro RNA-binding specificity.
The only known protein of hepatitis delta virus (HDV), the delta antigen, is found both within virus particles and within the nucleus of the infected cell, where it has one or more roles essential for RNA genome replication. Others have demonstrated that the antigen has the ability, in vitro, to specifically bind HDV RNA species. We report a further examination of this phenomenon, using partially purified recombinant protein, expressed as a fusion with the staphylococcal protein A. From Northwestern (RNA-immunoblot) analyses with both complete and various subdomains of HDV genomic and antigenomic RNAs, we found that a necessary feature for specific binding was that the RNA be able to fold to some extent into the so-called rodlike structure; this structure is a predicted intramolecular partial base-pairing of the circular RNA, with about 70% of all bases involved, so as to produce an unbranched rodlike structure. Six different subregions of the HDV rodlike structure, three on the genomic RNA and three on its complement, the antigenomic RNA, were tested and found to be sufficient for antigen binding. However, features in addition to the rodlike structure may also be necessary for specific binding, because we found that a similar structure present in the RNA of the potato spindle tuber viroid did not allow binding.
Chao
M
M
Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111.
Hsieh
S Y
SY
Taylor
J
J
D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism*; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000276:immunology*; D035923:Hepatitis delta Antigens; D015151:Immunoblotting; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015183:Restriction Mapping; D014772:Viroids / Q000235:genetics / Q000276:immunology
D000956:Antigens, Viral; D002352:Carrier Proteins; D035923:Hepatitis delta Antigens; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C442244:hepatitis delta virus large antigen