Poisson F, Roingeard P, Baillou A, Dubois F, Bonelli F, Calogero R A, Goudeau A
Départment de Microbiologie Médicale et Moléculaire, URA CNRS 1334, CHU Bretonneau, Tours, France.
J Gen Virol. 1993 Nov;74 ( Pt 11):2473-8. doi: 10.1099/0022-1317-74-11-2473.
Hepatitis delta antigen (HDAg), the only protein encoded by the hepatitis delta virus (HDV), binds specifically genomic and antigenomic strands of the HDV RNA. In a previous study, three recombinant HDAg subdomains were synthesized, covering residues 11 to 78, 79 to 163 and 164 to 212, and only the middle domain was shown to be responsible for the binding to HDV RNA. To investigate HDAg sequences involved in HDV RNA binding, we synthesized five peptides, 15 to 29 residues in length, and tested their ability to bind HDV RNA using a simple non-radioactive ELISA with digoxigenin-labelled HDV genomic or antigenomic RNA probes. The specificity of interactions was demonstrated by comparison with control peptides and non-HDV RNA probes, and with an inhibition assay using recombinant HDAg. The HDAg-binding domain found within the middle region (79 to 163) of HDAg was more finely mapped: it is located between residues 79 and 107. In addition, another domain (residues 2 to 27) of HDAg was also found to bind specifically to HDV RNA. These two peptides share sequence similarities at residues 2 to 10 and 97 to 107 with other RNA-binding domains.
丁型肝炎抗原（HDAg）是丁型肝炎病毒（HDV）唯一编码的蛋白质，它能特异性结合HDV RNA的基因组链和反基因组链。在先前的一项研究中，合成了三个重组HDAg亚结构域，覆盖第11至78位、第79至163位和第164至212位残基，结果显示只有中间结构域负责与HDV RNA结合。为了研究参与HDV RNA结合的HDAg序列，我们合成了5个长度为15至29个残基的肽段，并使用地高辛标记的HDV基因组或反基因组RNA探针通过简单的非放射性ELISA检测它们结合HDV RNA的能力。通过与对照肽段、非HDV RNA探针比较，以及使用重组HDAg进行抑制试验，证明了相互作用的特异性。在HDAg中间区域（79至163）内发现的HDAg结合结构域被更精确地定位：它位于第79至107位残基之间。此外，还发现HDAg的另一个结构域（第2至27位残基）也能特异性结合HDV RNA。这两个肽段在第2至10位和第97至107位残基处与其他RNA结合结构域具有序列相似性。
