An avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration

Michael Katzman, R. A. Katz, A. M. Skalka, J. Leis

Research output: Contribution to journalArticle

219 Citations (Scopus)

Abstract

The purified integration protein (IN) of avian myeloblastosis virus is shown to nick double-stranded oligodeoxynucleotide substrates that mimic the ends of the linear form of viral DNA. In the presence of Mg2+, nicks are created 2 nucleotides from the 3' OH ends of both the U5 plus strand and the U3 minus strand. Similar cleavage is observed in the presence of Mn2+ but only when the extent of the reaction is limited. Neither the complementary strands nor sequences representing the termini of human immunodeficiency virus type 1 DNA were cleaved at analogous positions. Analysis of a series of substrates containing U5 base substitutions has defined the sequence requirements for site-selective nicking; nucleotides near the cleavage site are most critical for activity. The minimum substrate size required to demonstrate significant activity corresponds to the nearly perfect 15-base terminal inverted repeat. This in vitro activity of IN thus produces viral DNA ends that are joined to host DNA in vivo and corresponds to an expected early step in the integrative recombination reaction. These results provide the first enzymatic support using purified retroviral proteins for a linear DNA precursor to the integrated provirus.

Original languageEnglish (US)
Pages (from-to)5319-5327
Number of pages9
JournalJournal of virology
Volume63
Issue number12
StatePublished - Jan 1 1989

Fingerprint

Viral DNA
DNA
Nucleotides
Avian Myeloblastosis Virus
Proviruses
Proteins
proteins
Terminal Repeat Sequences
Oligodeoxyribonucleotides
Avian myeloblastosis virus
Genetic Recombination
nucleotides
HIV-1
proviruses
terminal repeat sequences
Human immunodeficiency virus 1
avian retrovirus proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

@article{cd71ff70d59b49e498c3b6ede57358ab,
title = "An avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration",
abstract = "The purified integration protein (IN) of avian myeloblastosis virus is shown to nick double-stranded oligodeoxynucleotide substrates that mimic the ends of the linear form of viral DNA. In the presence of Mg2+, nicks are created 2 nucleotides from the 3' OH ends of both the U5 plus strand and the U3 minus strand. Similar cleavage is observed in the presence of Mn2+ but only when the extent of the reaction is limited. Neither the complementary strands nor sequences representing the termini of human immunodeficiency virus type 1 DNA were cleaved at analogous positions. Analysis of a series of substrates containing U5 base substitutions has defined the sequence requirements for site-selective nicking; nucleotides near the cleavage site are most critical for activity. The minimum substrate size required to demonstrate significant activity corresponds to the nearly perfect 15-base terminal inverted repeat. This in vitro activity of IN thus produces viral DNA ends that are joined to host DNA in vivo and corresponds to an expected early step in the integrative recombination reaction. These results provide the first enzymatic support using purified retroviral proteins for a linear DNA precursor to the integrated provirus.",
author = "Michael Katzman and Katz, {R. A.} and Skalka, {A. M.} and J. Leis",
year = "1989",
month = "1",
day = "1",
language = "English (US)",
volume = "63",
pages = "5319--5327",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "12",

}

An avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration. / Katzman, Michael; Katz, R. A.; Skalka, A. M.; Leis, J.

In: Journal of virology, Vol. 63, No. 12, 01.01.1989, p. 5319-5327.

Research output: Contribution to journalArticle

TY - JOUR

T1 - An avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration

AU - Katzman, Michael

AU - Katz, R. A.

AU - Skalka, A. M.

AU - Leis, J.

PY - 1989/1/1

Y1 - 1989/1/1

N2 - The purified integration protein (IN) of avian myeloblastosis virus is shown to nick double-stranded oligodeoxynucleotide substrates that mimic the ends of the linear form of viral DNA. In the presence of Mg2+, nicks are created 2 nucleotides from the 3' OH ends of both the U5 plus strand and the U3 minus strand. Similar cleavage is observed in the presence of Mn2+ but only when the extent of the reaction is limited. Neither the complementary strands nor sequences representing the termini of human immunodeficiency virus type 1 DNA were cleaved at analogous positions. Analysis of a series of substrates containing U5 base substitutions has defined the sequence requirements for site-selective nicking; nucleotides near the cleavage site are most critical for activity. The minimum substrate size required to demonstrate significant activity corresponds to the nearly perfect 15-base terminal inverted repeat. This in vitro activity of IN thus produces viral DNA ends that are joined to host DNA in vivo and corresponds to an expected early step in the integrative recombination reaction. These results provide the first enzymatic support using purified retroviral proteins for a linear DNA precursor to the integrated provirus.

AB - The purified integration protein (IN) of avian myeloblastosis virus is shown to nick double-stranded oligodeoxynucleotide substrates that mimic the ends of the linear form of viral DNA. In the presence of Mg2+, nicks are created 2 nucleotides from the 3' OH ends of both the U5 plus strand and the U3 minus strand. Similar cleavage is observed in the presence of Mn2+ but only when the extent of the reaction is limited. Neither the complementary strands nor sequences representing the termini of human immunodeficiency virus type 1 DNA were cleaved at analogous positions. Analysis of a series of substrates containing U5 base substitutions has defined the sequence requirements for site-selective nicking; nucleotides near the cleavage site are most critical for activity. The minimum substrate size required to demonstrate significant activity corresponds to the nearly perfect 15-base terminal inverted repeat. This in vitro activity of IN thus produces viral DNA ends that are joined to host DNA in vivo and corresponds to an expected early step in the integrative recombination reaction. These results provide the first enzymatic support using purified retroviral proteins for a linear DNA precursor to the integrated provirus.

UR - http://www.scopus.com/inward/record.url?scp=0024431589&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024431589&partnerID=8YFLogxK

M3 - Article

C2 - 2555556

AN - SCOPUS:0024431589

VL - 63

SP - 5319

EP - 5327

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 12

ER -