IN-VITRO ACTIVITIES OF PURIFIED VISNA VIRUS INTEGRASE

被引:30
作者
KATZMAN, M [1 ]
SUDOL, M [1 ]
机构
[1] PENN STATE UNIV,COLL MED,MILTON S HERSHEY MED CTR,DEPT MICROBIOL & IMMUNOL,HERSHEY,PA 17033
关键词
D O I
10.1128/JVI.68.6.3558-3569.1994
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:3558 / 3569
页数:12
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