Multivalent interactions essential for lentiviral integrase function - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Nature Communications Année : 2022

Multivalent interactions essential for lentiviral integrase function

Multivalent interactions essential for lentiviral integrase function.

Vidya Chivukula
Dominika Gruszka
Zelin Shan
Parmit Singh
Valerie Pye
Rebecca Mclean
Gregory Bedwell
  • Fonction : Auteur
Andrea Nans
Nicola Cook
Hind Fadel
  • Fonction : Auteur
Eric Poeschla
Dmitry Lyumkis
Hasan Yardimci
Alan Engelman
Peter Cherepanov

Résumé

Abstract A multimer of retroviral integrase (IN) synapses viral DNA ends within a stable intasome nucleoprotein complex for integration into a host cell genome. Reconstitution of the intasome from the maedi-visna virus (MVV), an ovine lentivirus, revealed a large assembly containing sixteen IN subunits 1 . Herein, we report cryo-EM structures of the lentiviral intasome prior to engagement of target DNA and following strand transfer, refined at 3.4 and 3.5 Å resolution, respectively. The structures elucidate details of the protein-protein and protein-DNA interfaces involved in lentiviral intasome formation. We show that the homomeric interfaces involved in IN hexadecamer formation and the α-helical configuration of the linker connecting the C-terminal and catalytic core domains are critical for MVV IN strand transfer activity in vitro and for virus infectivity. Single-molecule microscopy in conjunction with photobleaching reveals that the MVV intasome can bind a variable number, up to sixteen molecules, of the lentivirus-specific host factor LEDGF/p75. Concordantly, ablation of endogenous LEDGF/p75 results in gross redistribution of MVV integration sites in human and ovine cells. Our data confirm the importance of the expanded architecture observed in cryo-EM studies of lentiviral intasomes and suggest that this organization underlies multivalent interactions with chromatin for integration targeting to active genes.

Dates et versions

hal-04246036 , version 1 (17-10-2023)

Identifiants

Citer

Allison Ballandras-Colas, Vidya Chivukula, Dominika Gruszka, Zelin Shan, Parmit Singh, et al.. Multivalent interactions essential for lentiviral integrase function. Nature Communications, 2022, 13 (1), pp.2416. ⟨10.1038/s41467-022-29928-8⟩. ⟨hal-04246036⟩
9 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More