HIV-1 integrase crosslinked oligomers are active in vitro

被引：154

作者：

Faure, A ^{[1
]}

Calmels, C ^{[1
]}

Desjobert, C ^{[1
]}

Castroviejo, M ^{[1
]}

Caumont-Sarcos, A ^{[1
]}

Tarrago-Litvak, L ^{[1
]}

Litvak, S ^{[1
]}

Parissi, V ^{[1
]}

机构：

[1] Univ Bordeaux 2, CNRS, UMR 5097, IFR 66 Pathol Infect & Canc, F-33076 Bordeaux, France

来源：

NUCLEIC ACIDS RESEARCH | 2005年 / 33卷 / 03期

关键词：

D O I：

10.1093/nar/gki241

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The oligomeric state of active human immunodeficiency virus type 1 (HIV-1) integrase (IN) has not been clearly elucidated. We analyzed the activity of the different purified oligomeric forms of recombinant IN obtained after stabilization by platinum crosslinking. The crosslinked tetramer isolated by gel chromatography was able to catalyze the full-site integration of the two viral LTR ends into a target DNA in vitro, whereas the isolated dimeric form of the enzyme was involved in the processing and integration of only one viral end. Accurate concerted integration by IN tetramers was confirmed by cloning and sequencing. Kinetic studies of DNA-integrase complexes led us to propose a model explaining the formation of an active complex. Our data suggest that the tetrameric IN bound to the viral DNA ends is the minimal complex involved in the concerted integration of both LTRs and should be the oligomeric form targeted by future inhibitors.

引用

页码：977 / 986

页数：10

共 41 条

[11] Three-dimensional structure of the Tn5 synaptic complex transposition intermediate [J].

Davies, DR ;

Goryshin, IY ;

Reznikoff, WS ;

Rayment, I .

SCIENCE, 2000, 289 (5476) :77-85

[12] Oligomeric states of the HIV-1 integrase as measured by time-resolved fluorescence anisotropy [J].

Deprez, E ;

Tauc, P ;

Leh, H ;

Mouscadet, JF ;

Auclair, C ;

Brochon, JC .

BIOCHEMISTRY, 2000, 39 (31) :9275-9284

[13] DNA binding induces dissociation of the multimeric form of HIV-1 integrase: A time-resolved fluorescence anisotropy study [J].

Deprez, E ;

Tauc, P ;

Leh, H ;

Mouscadet, JF ;

Auclair, C ;

Hawkins, ME ;

Brochon, JC .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (18) :10090-10095

[14] p66/p51 and p51/p51 recombinant forms of reverse transcriptase from human immunodeficiency virus type 1 -: Interactions with primer tRNALys3, initiation of cDNA synthesis, and effect of inhibitors [J].

Dufour, E ;

El Dirani-Diab, R ;

Boulmé, F ;

Fournier, M ;

Nevinsky, G ;

Tarrago-Litvak, L ;

Litvak, S ;

Andreola, ML .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1998, 251 (1-2) :487-495

[15] A STABLE COMPLEX BETWEEN INTEGRASE AND VIRAL-DNA ENDS MEDIATES HUMAN-IMMUNODEFICIENCY-VIRUS INTEGRATION IN-VITRO [J].

ELLISON, V ;

BROWN, PO .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (15) :7316-7320

[16] IDENTIFICATION OF DISCRETE FUNCTIONAL DOMAINS OF HIV-1 INTEGRASE AND THEIR ORGANIZATION WITHIN AN ACTIVE MULTIMERIC COMPLEX [J].

ENGELMAN, A ;

BUSHMAN, FD ;

CRAIGIE, R .

EMBO JOURNAL, 1993, 12 (08) :3269-3275

[17] HIV-1 integrase inhibitors that compete with the target DNA substrate define a unique strand transfer conformation for integrase [J].

Espeseth, AS ;

Felock, P ;

Wolfe, A ;

Witmer, M ;

Grobler, J ;

Anthony, N ;

Egbertson, M ;

Melamed, JY ;

Young, S ;

Hamill, T ;

Cole, JL ;

Hazuda, DJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (21) :11244-11249

[18] Human immunodeficiency virus type 1 integrase: arrangement of protein domains in active cDNA complexes [J].

Gao, K ;

Butler, SL ;

Bushman, F .

EMBO JOURNAL, 2001, 20 (13) :3565-3576

[19] CONCERTED INTEGRATION OF RETROVIRUS-LIKE DNA BY HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 INTEGRASE [J].

GOODARZI, G ;

IM, GJ ;

BRACKMANN, K ;

GRANDGENETT, D .

JOURNAL OF VIROLOGY, 1995, 69 (10) :6090-6097

[20] STUDIES ON TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS [J].

HANAHAN, D .

JOURNAL OF MOLECULAR BIOLOGY, 1983, 166 (04) :557-580

← 1 2 3 4 5 →