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NHEJ pathway is involved in post-integrational DNA repair due to Ku70 binding to HIV-1 integrase.

Abstract：

BACKGROUND:HIV-1 integration results in genomic DNA gaps that are repaired by cellular DNA repair pathways. This step of the lentiviral life cycle remains poorly understood despite its crucial importance for successful replication. We and others reported that Ku70 protein of the non-homologous end joining pathway (NHEJ) directly binds HIV-1 integrase (IN). Here, we studied the importance of this interaction for post-integrational gap repair and the recruitment of NHEJ factors in this process. RESULTS:We engineered HIV-based pseudovirus with mutant IN defective in Ku70 binding and generated heterozygous Ku70, Ku80 and DNA-PKcs human knockout (KO) cells using CRISPR/Cas9. KO of either of these proteins or inhibition of DNA-PKcs catalytic activity substantially decreased the infectivity of HIV-1 with native IN but not with the mutant one. We used a recently developed qPCR assay for the measurement of gap repair efficiency to show that HIV-1 with mutant IN was defective in DNA post-integrational repair, whereas the wild type virus displayed such a defect only when NHEJ system was disrupted in any way. This effect was present in CRISPR/Cas9 modified 293T cells, in Jurkat and CEM lymphoid lines and in primary human PBMCs. CONCLUSIONS:Our data provide evidence that IN recruits DNA-PK to the site of HIV-1 post-integrational repair due to Ku70 binding-a novel finding that explains the involvement of DNA-PK despite the absence of free double stranded DNA breaks. In addition, our data clearly indicate the importance of interactions between HIV-1 IN and Ku70 in HIV-1 replication at the post-integrational repair step.

journal_name

Retrovirology

journal_title

Retrovirology

authors

Knyazhanskaya E,Anisenko A,Shadrina O,Kalinina A,Zatsepin T,Zalevsky A,Mazurov D,Gottikh M

doi

10.1186/s12977-019-0492-z

subject

Has Abstract

pub_date

2019-11-06 00:00:00

pages

30

issue

1

issn

1742-4690

pii

10.1186/s12977-019-0492-z

journal_volume

16

pub_type

杂志文章

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