Hepatitis delta antigen binds to the clamp of RNA polymerase II and affects transcriptional fidelity.
Hepatitis delta virus (HDV) is an RNA virus whose replication and transcription are considered to proceed via RNA-dependent RNA synthesis by RNA polymerase II (Pol II), and the viral protein called hepatitis delta antigen (HDAg) is essential for these processes. HDAg was previously shown to stimulate Pol II elongation on both DNA and RNA templates in vitro. Here, the mechanism of elongation control by HDAg was investigated because it serves as a prototype of cellular transcription elongation factors and also plays an interesting role in HDV proliferation. With site-specific photocrosslinking and transcription using reconstituted elongation complexes, evidence is presented that HDAg functionally interacts with the clamp of Pol II, a mobile structure that holds DNA and RNA in place. Strikingly, HDAg not only increases the rate of elongation but also affects the decision of which nucleotide is incorporated. These and our previous findings lead us to propose a model in which HDAg interacts with and loosens the clamp, and thereby accelerates forward translocation of Pol II at the cost of fidelity. By reducing transcriptional fidelity in terms of not only discrimination of incoming nucleotides but also recognition of templates, HDAg may facilitate the unusual RNA-dependent RNA synthesis by Pol II.
Yamaguchi
Yuki
Y
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226-8501, Japan.
Mura
Takashi
T
Chanarat
Sittinan
S
Okamoto
Sachiko
S
Handa
Hiroshi
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D017124:Conserved Sequence; D019143:Evolution, Molecular; D006367:HeLa Cells; D035923:Hepatitis delta Antigens / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012319:RNA Polymerase II / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D014158:Transcription, Genetic
D035923:Hepatitis delta Antigens; C500816:POLR2B protein, human; D012319:RNA Polymerase II; C410949:RNA polymerase II largest subunit