Four conserved cysteine residues of the hepatitis B virus polymerase are critical for RNA pregenome encapsidation

Seahee Kim, Jehan Lee, Wang-Shick Ryu

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Hepadnaviruses replicate via reverse transcription of an RNA template, the pregenomic RNA (pgRNA). Although hepadnaviral polymerase (Pol) and retroviral reverse transcriptase are distantly related, some of their features are distinct. In particular, Pol contains two additional N-terminal subdomains, the terminal protein and spacer subdomains. Since much of the spacer subdomain can be deleted without detrimental effects to hepatitis B virus (HBV) replication, this subdomain was previously thought to serve only as a spacer that links the terminal protein and reverse transcriptase subdomains. Unexpectedly, we found that the C terminus of the spacer subdomain is indispensable for the encapsidation of pgRNA. Alanine-scanning mutagenesis revealed that four conserved cysteine residues, three at the C terminus of the spacer subdomain and one at the N terminus of the reverse transcriptase subdomain, are critical for encapsidation. The inability of the mutant Pol proteins to incorporate into nucleocapsid particles, together with other evidence, argued that the four conserved cysteine residues are critical for RNA binding. One implication is that these four cysteine residues might form a putative zinc finger motif. Based on these findings, we speculate that the RNA binding activity of HBV Pol may be mediated by this newly identified putative zinc finger motif.

Original languageEnglish
Pages (from-to)8032-8040
Number of pages9
JournalJournal of Virology
Volume83
Issue number16
DOIs
Publication statusPublished - 2009 Aug 1

Fingerprint

Hepatitis B virus
DNA-Directed RNA Polymerases
Cysteine
cysteine
RNA
RNA-directed DNA polymerase
RNA-Directed DNA Polymerase
zinc finger motif
Zinc Fingers
Hepadnaviridae
Nucleocapsid
nucleocapsid
reverse transcription
proteins
Mutant Proteins
Virus Replication
virus replication
mutagenesis
Mutagenesis
Alanine

All Science Journal Classification (ASJC) codes

  • Immunology
  • Virology

Cite this

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Four conserved cysteine residues of the hepatitis B virus polymerase are critical for RNA pregenome encapsidation. / Kim, Seahee; Lee, Jehan; Ryu, Wang-Shick.

In: Journal of Virology, Vol. 83, No. 16, 01.08.2009, p. 8032-8040.

Research output: Contribution to journalArticle

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