Distinct requirement for two stages of protein-primed initiation of reverse transcription in hepadnaviruses

Xingtai Wang, Jianming Hu

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Reverse transcription in hepadnaviruses is primed by the viral reverse transcriptase (RT) (protein priming) and requires the specific interaction between the RT and a viral RNA signal termed ε, which bears the specific template sequence for protein priming. The product of protein priming is a short oligodeoxynucleotide which represents the 5′ end of the viral minus-strand DNA and is covalently attached to the RT. We have now identified truncated RT variants from the duck hepatitis B virus that were fully active in the initial step of protein priming, i.e., the covalent attachment of the first nucleotide to the protein (RT deoxynucleotidylation), but defective in any subsequent DNA polymerization. A short sequence in the RT domain was localized that was dispensable for RT deoxynucleotidylation but essential for the subsequent DNA polymerization. These results have thus revealed two distinct stages of protein priming, i.e., the initial attachment of the first nucleotide to the RT (RT deoxynucleotidylation or initiation of protein priming) and the subsequent DNA synthesis (polymerization) to complete protein priming, with the second step entailing additional RT sequences. Two models are proposed to explain the observed differential sequence requirement for the two distinct stages of the protein priming reaction.

Original languageEnglish (US)
Pages (from-to)5857-5865
Number of pages9
JournalJournal of virology
Volume76
Issue number12
DOIs
StatePublished - Jun 5 2002

Fingerprint

Hepadnaviridae
reverse transcription
RNA-directed DNA polymerase
RNA-Directed DNA Polymerase
Reverse Transcription
Proteins
proteins
Polymerization
polymerization
DNA
Duck hepatitis B virus
Nucleotides
Duck Hepatitis B Viruses
nucleotides
Oligodeoxyribonucleotides
protein products
Viral RNA

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

@article{62d69d6fe7034d0fbe3b054e1ccf139b,
title = "Distinct requirement for two stages of protein-primed initiation of reverse transcription in hepadnaviruses",
abstract = "Reverse transcription in hepadnaviruses is primed by the viral reverse transcriptase (RT) (protein priming) and requires the specific interaction between the RT and a viral RNA signal termed ε, which bears the specific template sequence for protein priming. The product of protein priming is a short oligodeoxynucleotide which represents the 5′ end of the viral minus-strand DNA and is covalently attached to the RT. We have now identified truncated RT variants from the duck hepatitis B virus that were fully active in the initial step of protein priming, i.e., the covalent attachment of the first nucleotide to the protein (RT deoxynucleotidylation), but defective in any subsequent DNA polymerization. A short sequence in the RT domain was localized that was dispensable for RT deoxynucleotidylation but essential for the subsequent DNA polymerization. These results have thus revealed two distinct stages of protein priming, i.e., the initial attachment of the first nucleotide to the RT (RT deoxynucleotidylation or initiation of protein priming) and the subsequent DNA synthesis (polymerization) to complete protein priming, with the second step entailing additional RT sequences. Two models are proposed to explain the observed differential sequence requirement for the two distinct stages of the protein priming reaction.",
author = "Xingtai Wang and Jianming Hu",
year = "2002",
month = "6",
day = "5",
doi = "10.1128/JVI.76.12.5857-5865.2002",
language = "English (US)",
volume = "76",
pages = "5857--5865",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "12",

}

Distinct requirement for two stages of protein-primed initiation of reverse transcription in hepadnaviruses. / Wang, Xingtai; Hu, Jianming.

In: Journal of virology, Vol. 76, No. 12, 05.06.2002, p. 5857-5865.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Distinct requirement for two stages of protein-primed initiation of reverse transcription in hepadnaviruses

AU - Wang, Xingtai

AU - Hu, Jianming

PY - 2002/6/5

Y1 - 2002/6/5

N2 - Reverse transcription in hepadnaviruses is primed by the viral reverse transcriptase (RT) (protein priming) and requires the specific interaction between the RT and a viral RNA signal termed ε, which bears the specific template sequence for protein priming. The product of protein priming is a short oligodeoxynucleotide which represents the 5′ end of the viral minus-strand DNA and is covalently attached to the RT. We have now identified truncated RT variants from the duck hepatitis B virus that were fully active in the initial step of protein priming, i.e., the covalent attachment of the first nucleotide to the protein (RT deoxynucleotidylation), but defective in any subsequent DNA polymerization. A short sequence in the RT domain was localized that was dispensable for RT deoxynucleotidylation but essential for the subsequent DNA polymerization. These results have thus revealed two distinct stages of protein priming, i.e., the initial attachment of the first nucleotide to the RT (RT deoxynucleotidylation or initiation of protein priming) and the subsequent DNA synthesis (polymerization) to complete protein priming, with the second step entailing additional RT sequences. Two models are proposed to explain the observed differential sequence requirement for the two distinct stages of the protein priming reaction.

AB - Reverse transcription in hepadnaviruses is primed by the viral reverse transcriptase (RT) (protein priming) and requires the specific interaction between the RT and a viral RNA signal termed ε, which bears the specific template sequence for protein priming. The product of protein priming is a short oligodeoxynucleotide which represents the 5′ end of the viral minus-strand DNA and is covalently attached to the RT. We have now identified truncated RT variants from the duck hepatitis B virus that were fully active in the initial step of protein priming, i.e., the covalent attachment of the first nucleotide to the protein (RT deoxynucleotidylation), but defective in any subsequent DNA polymerization. A short sequence in the RT domain was localized that was dispensable for RT deoxynucleotidylation but essential for the subsequent DNA polymerization. These results have thus revealed two distinct stages of protein priming, i.e., the initial attachment of the first nucleotide to the RT (RT deoxynucleotidylation or initiation of protein priming) and the subsequent DNA synthesis (polymerization) to complete protein priming, with the second step entailing additional RT sequences. Two models are proposed to explain the observed differential sequence requirement for the two distinct stages of the protein priming reaction.

UR - http://www.scopus.com/inward/record.url?scp=0036111150&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036111150&partnerID=8YFLogxK

U2 - 10.1128/JVI.76.12.5857-5865.2002

DO - 10.1128/JVI.76.12.5857-5865.2002

M3 - Article

C2 - 12021318

AN - SCOPUS:0036111150

VL - 76

SP - 5857

EP - 5865

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 12

ER -