Abstract:
:The Moloney murine leukemia virus (MLV) is a prototype gammaretrovirus requiring nuclear disassembly before DNA integration. In the nucleus, integration site selection towards promoter/enhancer elements is mediated by the host factor bromo- and extraterminal domain (BET) proteins (bromodomain (Brd) proteins 2, 3 and 4). MLV-based retroviral vectors are used in gene therapy trials. In some trials leukemia occurred through integration of the MLV vector in close proximity to cellular oncogenes. BET-mediated integration is poorly understood and the nature of integrase oligomers heavily debated. Here, we created wild-type infectious MLV vectors natively incorporating fluorescent labeled IN and performed single-molecule intensity and Förster resonance energy transfer experiments. The nuclear localization of the MLV pre-integration complex neither altered the IN content, nor its quaternary structure. Instead, BET-mediated interaction of the MLV intasome with chromatin in the post-mitotic nucleus reshaped its quaternary structure.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Borrenberghs D,Zurnic I,De Wit F,Acke A,Dirix L,Cereseto A,Debyser Z,Hendrix Jdoi
10.1093/nar/gky1157subject
Has Abstractpub_date
2019-02-20 00:00:00pages
1195-1210issue
3eissn
0305-1048issn
1362-4962pii
5184726journal_volume
47pub_type
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