Multivalent interactions essential for lentiviral integrase function
Springer Nature, 2022
Online
academicJournal
Zugriff:
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 subunits1. 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.
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Multivalent interactions essential for lentiviral integrase function
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Autor/in / Beteiligte Person: | Ballandras-Colas, A ; Chivukula, V ; Gruszka, D ; Shan, Z ; Singh, P ; Pye, V ; McLean, R ; Bedwell, G ; Li, W ; Nans, A ; Cook, N ; Fadel, H ; Poeschla, E ; Griffiths, D ; Vargas, J ; Taylor, I ; Lyumkis, D ; Yardimci, H ; Engelman, A ; Cherepanov, P |
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Veröffentlichung: | Springer Nature, 2022 |
Medientyp: | academicJournal |
DOI: | 10.1038/s41467-022-29928-8 |
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