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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
7
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pubmed:dateCreated |
2002-3-8
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pubmed:abstractText |
Replication of retroviruses requires integration of the linear viral DNA genome into the host chromosomes. Integration requires the viral integrase (IN), located in high-molecular-weight nucleoprotein complexes termed preintegration complexes (PIC). The PIC inserts the two viral DNA termini in a concerted manner into chromosomes in vivo as well as exogenous target DNA in vitro. We reconstituted nucleoprotein complexes capable of efficient concerted (full-site) integration using recombinant wild-type human immunodeficiency virus type I (HIV-1) IN with linear retrovirus-like donor DNA (480 bp). In addition, no cellular or viral protein cofactors are necessary for purified bacterial recombinant HIV-1 IN to mediate efficient full-site integration of two donor termini into supercoiled target DNA. At about 30 nM IN (20 min at 37 degrees C), approximately 15 and 8% of the input donor is incorporated into target DNA, producing half-site (insertion of one viral DNA end per target) and full-site integration products, respectively. Sequencing the donor-target junctions of full-site recombinants confirms that 5-bp host site duplications have occurred with a fidelity of about 70%, similar to the fidelity when using IN derived from nonionic detergent lysates of HIV-1 virions. A key factor allowing recombinant wild-type HIV-1 IN to mediate full-site integration appears to be the avoidance of high IN concentrations in its purification (about 125 microg/ml) and in the integration assay (<50 nM). The results show that recombinant HIV-1 IN may not be significantly defective for full-site integration. The findings further suggest that a high concentration or possibly aggregation of IN is detrimental to the assembly of correct nucleoprotein complexes for full-site integration.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10074149,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10187830,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10219094,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10233971,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10358097,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10388663,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10400764,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10482559,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10516007,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10924120,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10924121,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-10949691,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-11152486,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-11264345,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-11346660,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-1328665,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-1404595,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-1847499,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-2002549,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-2164223,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-2167180,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-2170022,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-2562907,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-3032450,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-7666512,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-7971276,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-848760,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-8631811,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-8648691,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-8942990,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-8993331,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9037048,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9038339,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9168883,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9188609,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9295344,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9405379,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9499068,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9578550,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9733892,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11884535-9790878
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0022-538X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
76
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
3105-13
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
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pubmed:year |
2002
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pubmed:articleTitle |
Efficient concerted integration by recombinant human immunodeficiency virus type 1 integrase without cellular or viral cofactors.
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pubmed:affiliation |
Institute for Molecular Virology, St. Louis University Health Sciences Center, St. Louis, Missouri 63110, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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