LEDGF/p75 functions downstream from preintegration complex formation to effect gene-specific HIV-1 integration

被引:371
作者
Shun, Ming-Chieh
Raghavendra, Nidhanapati K.
Vandegraaff, Nick
Daigle, Janet E.
Hughes, Siobhan
Kellam, Paul
Cherepanov, Peter [1 ]
Engelman, Alan
机构
[1] Harvard Univ, Sch Med, Dana Farber Canc Inst,Div AIDS, Dept Canc Immunol & AIDS, Boston, MA 02115 USA
[2] Imperial Coll London, Div Med, London W2 1PG, England
[3] UCL, Dept Infect, London W1T 4JF, England
基金
英国医学研究理事会;
关键词
integrase; LEDGF/p75; HIV-1; AIDS; transcription; integration;
D O I
10.1101/gad.1565107
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
LEDGF/p75 directly interacts with lentiviral integrase proteins and can modulate their enzymatic activities and chromosomal association. A novel genetic knockout model was established that allowed us for the first time to analyze HIV-1 integration in the absence of LEDGF/p75 protein. Supporting a crucial role for the cofactor in viral replication, HIV-1 vector integration and reporter gene expression were significantly reduced in LEDGF-null cells. Yet, integrase processed the viral cDNA termini normally and maintained its local target DNA sequence preference during integration. Preintegration complexes extracted from knockout cells moreover supported normal levels of DNA strand transfer activity in vitro. In contrast, HIV-1 lost its strong bias toward integrating into transcription units, displaying instead increased affinity for promoter regions and CpG islands. Our results reveal LEDGF/p75 as a critical targeting factor, commandeering lentiviruses from promoter- and/or CpG island-proximal pathways that are favored by other members of Retroviridae. Akin to yeast retrotransposons, disrupting the lentiviral targeting mechanism significantly perturbs overall integration.
引用
收藏
页码:1767 / 1778
页数:12
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