In vitro epsilon RNA-dependent protein priming activity of human hepatitis B virus polymerase

Scott A. Jones, Rajeev Boregowd, Thomas E. Spratt, Jianming Hu

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Hepatitis B virus (HBV) replicates its DNA genome through reverse transcription of a pregenomic RNA (pgRNA) by using a multifunctional polymerase (HP). A critical function of HP is its specific recognition of a viral RNA signal termed ε (Hε) located on pgRNA, which is required for specific packaging of pgRNA into viral nucleocapsids and initiation of viral reverse transcription. HP initiates reverse transcription by using itself as a protein primer (protein priming) and Hε as the obligatory template. We have purified HP from human cells that retained Hε binding activity in vitro. Furthermore, HP purified as a complex with Hε, but not HP alone, displayed in vitro protein priming activity. While the HP-Hε interaction in vitro and in vivo required the Hε internal bulge, but not its apical loop, and was not significantly affected by the cap-Hε distance, protein priming required both the Hε apical loop and internal bulge, as well as a short distance between the cap and Hε, mirroring the requirements for RNA packaging. These studies have thus established new HBV protein priming and RNA binding assays that should greatly facilitate the dissection of the requirements and molecular mechanisms of HP-Hε interactions, RNA packaging, and protein priming.

Original languageEnglish (US)
Pages (from-to)5134-5150
Number of pages17
JournalJournal of virology
Volume86
Issue number9
DOIs
StatePublished - May 1 2012

Fingerprint

Hepatitis B virus
Human Activities
RNA
Product Packaging
reverse transcription
Reverse Transcription
Viral RNA
Proteins
proteins
Nucleocapsid
RNA-Binding Proteins
nucleocapsid
Dissection
In Vitro Techniques
Genome
DNA
genome
assays
RNA packaging
cells

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

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In vitro epsilon RNA-dependent protein priming activity of human hepatitis B virus polymerase. / Jones, Scott A.; Boregowd, Rajeev; Spratt, Thomas E.; Hu, Jianming.

In: Journal of virology, Vol. 86, No. 9, 01.05.2012, p. 5134-5150.

Research output: Contribution to journalArticle

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