Structural basis for HIV-1 DNA integration in the human genome, role of the LEDGF/P75 cofactor.
Integration of the human immunodeficiency virus (HIV-1) cDNA into the human genome is catalysed by integrase. Several studies have shown the importance of the interaction of cellular cofactors with integrase for viral integration and infectivity. In this study, we produced a stable and functional complex between the wild-type full-length integrase (IN) and the cellular cofactor LEDGF/p75 that shows enhanced in vitro integration activity compared with the integrase alone. Mass spectrometry analysis and the fitting of known atomic structures in cryo negatively stain electron microscopy (EM) maps revealed that the functional unit comprises two asymmetric integrase dimers and two LEDGF/p75 molecules. In the presence of DNA, EM revealed the DNA-binding sites and indicated that, in each asymmetric dimer, one integrase molecule performs the catalytic reaction, whereas the other one positions the viral DNA in the active site of the opposite dimer. The positions of the target and viral DNAs for the 3' processing and integration reaction shed light on the integration mechanism, a process with wide implications for the understanding of viral-induced pathologies.
Michel
Fabrice
F
IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire), Département de Biologie et de Génomique Structurales, UDS, CNRS, INSERM, Illkirch, France.
Crucifix
Corinne
C
Granger
Florence
F
Eiler
Sylvia
S
Mouscadet
Jean-François
JF
Korolev
Sergei
S
Agapkina
Julia
J
Ziganshin
Rustam
R
Gottikh
Marina
M
Nazabal
Alexis
A
Emiliani
Stéphane
S
Benarous
Richard
R
Moras
Dino
D
Schultz
Patrick
P
Ruff
Marc
M
D020285:Cryoelectron Microscopy; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015894:Genome, Human; D019427:HIV Integrase / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry; D008958:Models, Molecular; D011487:Protein Conformation; D016662:Virus Integration; D014779:Virus Replication
D004279:DNA, Viral; D036341:Intercellular Signaling Peptides and Proteins; C119380:lens epithelium-derived growth factor; D019427:HIV Integrase