In vitro activities of purified visna virus integrase

Michael Katzman, Malgorzata Sudol

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Although integration generally is considered a critical step in the retrovirus life cycle, it has been reported that visna virus, which causes degenerative neurologic disease in sheep, can productively infect sheep choroid plexus cells without detectable integration. To ascertain whether the integrase (IN) of visna virus is an inherently defective enzyme and to create tools for further study of integration of the phylogenetically related human immunodeficiency virus type 1 (HIV-1), we purified visna virus IN by using a bacterial expression system and applied various in vitro oligonucleotide- based assays to studying this protein. We found that visna virus IN demonstrates the full repertoire of in vitro functions characteristic of retroviral integrases. In particular, visna virus IN exhibits site-specific endonuclease activity following the invariant CA found two nucleotides from the 3' ends of viral DNA (processing activity), joins processed oligonucleotides to various sites on other oligonucleotides (strand transfer or integration activity), and reverses the integration reaction by resolving a complex that mimics one end of viral DNA integrated into host DNA (disintegration activity). In addition, although it has been reported that purified HIV-1 IN cannot specifically nick visna virus DNA ends, purified visna virus IN does specifically process and integrate HIV-1 DNA ends.

Original languageEnglish (US)
Pages (from-to)3558-3569
Number of pages12
JournalJournal of Virology
Volume68
Issue number6
StatePublished - Jun 1 1994

Fingerprint

Visna-maedi virus
Visna maedi virus
Integrases
oligonucleotides
Human immunodeficiency virus 1
Oligonucleotides
HIV-1
Sheep
Retroviridae
choroid plexus
sheep
Choroid Plexus
nervous system diseases
Endonucleases
In Vitro Techniques
Life Cycle Stages
Neurodegenerative Diseases
life cycle (organisms)
Nucleotides
nucleotides

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Katzman, Michael ; Sudol, Malgorzata. / In vitro activities of purified visna virus integrase. In: Journal of Virology. 1994 ; Vol. 68, No. 6. pp. 3558-3569.
@article{8eada5facee24bdf93fd0db461372535,
title = "In vitro activities of purified visna virus integrase",
abstract = "Although integration generally is considered a critical step in the retrovirus life cycle, it has been reported that visna virus, which causes degenerative neurologic disease in sheep, can productively infect sheep choroid plexus cells without detectable integration. To ascertain whether the integrase (IN) of visna virus is an inherently defective enzyme and to create tools for further study of integration of the phylogenetically related human immunodeficiency virus type 1 (HIV-1), we purified visna virus IN by using a bacterial expression system and applied various in vitro oligonucleotide- based assays to studying this protein. We found that visna virus IN demonstrates the full repertoire of in vitro functions characteristic of retroviral integrases. In particular, visna virus IN exhibits site-specific endonuclease activity following the invariant CA found two nucleotides from the 3' ends of viral DNA (processing activity), joins processed oligonucleotides to various sites on other oligonucleotides (strand transfer or integration activity), and reverses the integration reaction by resolving a complex that mimics one end of viral DNA integrated into host DNA (disintegration activity). In addition, although it has been reported that purified HIV-1 IN cannot specifically nick visna virus DNA ends, purified visna virus IN does specifically process and integrate HIV-1 DNA ends.",
author = "Michael Katzman and Malgorzata Sudol",
year = "1994",
month = "6",
day = "1",
language = "English (US)",
volume = "68",
pages = "3558--3569",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "6",

}

In vitro activities of purified visna virus integrase. / Katzman, Michael; Sudol, Malgorzata.

In: Journal of Virology, Vol. 68, No. 6, 01.06.1994, p. 3558-3569.

Research output: Contribution to journalArticle

TY - JOUR

T1 - In vitro activities of purified visna virus integrase

AU - Katzman, Michael

AU - Sudol, Malgorzata

PY - 1994/6/1

Y1 - 1994/6/1

N2 - Although integration generally is considered a critical step in the retrovirus life cycle, it has been reported that visna virus, which causes degenerative neurologic disease in sheep, can productively infect sheep choroid plexus cells without detectable integration. To ascertain whether the integrase (IN) of visna virus is an inherently defective enzyme and to create tools for further study of integration of the phylogenetically related human immunodeficiency virus type 1 (HIV-1), we purified visna virus IN by using a bacterial expression system and applied various in vitro oligonucleotide- based assays to studying this protein. We found that visna virus IN demonstrates the full repertoire of in vitro functions characteristic of retroviral integrases. In particular, visna virus IN exhibits site-specific endonuclease activity following the invariant CA found two nucleotides from the 3' ends of viral DNA (processing activity), joins processed oligonucleotides to various sites on other oligonucleotides (strand transfer or integration activity), and reverses the integration reaction by resolving a complex that mimics one end of viral DNA integrated into host DNA (disintegration activity). In addition, although it has been reported that purified HIV-1 IN cannot specifically nick visna virus DNA ends, purified visna virus IN does specifically process and integrate HIV-1 DNA ends.

AB - Although integration generally is considered a critical step in the retrovirus life cycle, it has been reported that visna virus, which causes degenerative neurologic disease in sheep, can productively infect sheep choroid plexus cells without detectable integration. To ascertain whether the integrase (IN) of visna virus is an inherently defective enzyme and to create tools for further study of integration of the phylogenetically related human immunodeficiency virus type 1 (HIV-1), we purified visna virus IN by using a bacterial expression system and applied various in vitro oligonucleotide- based assays to studying this protein. We found that visna virus IN demonstrates the full repertoire of in vitro functions characteristic of retroviral integrases. In particular, visna virus IN exhibits site-specific endonuclease activity following the invariant CA found two nucleotides from the 3' ends of viral DNA (processing activity), joins processed oligonucleotides to various sites on other oligonucleotides (strand transfer or integration activity), and reverses the integration reaction by resolving a complex that mimics one end of viral DNA integrated into host DNA (disintegration activity). In addition, although it has been reported that purified HIV-1 IN cannot specifically nick visna virus DNA ends, purified visna virus IN does specifically process and integrate HIV-1 DNA ends.

UR - http://www.scopus.com/inward/record.url?scp=0028237180&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028237180&partnerID=8YFLogxK

M3 - Article

C2 - 8189495

AN - SCOPUS:0028237180

VL - 68

SP - 3558

EP - 3569

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 6

ER -