Figure 3.

Unintegrated lentivirus expression is restricted by the SMC5/6 complex in an SLF2-dependent but SLF1-independent manner

(A and B) IP-MS identifies SMC5/6 complex components as SLF2-interactors.

(A) Immunoprecipitated material from endogenous SLF2 IP in SLF2-KO and WT Jurkat cells was analyzed by mass spectrometry. Interactors displayed in (B) satisfy inclusion criteria of being (1) undetected in IP from SLF2-KO cells and (2) detected with ≥3 peptides in IP from WT cells. Interactors are displayed in descending order by number of unique peptides identified. Representative dataset (n = 2). Purple bars indicate SLF2/SMC6 interactors identified by (Räschle et al., 2015)

(C–F) Knockout of SMC5/6 complex but not SLF1 increases unintegrated virus expression. Unintegrated virus reporter infection of mixed KO populations 7 days post-sgRNA transduction of Cas9-Jurkat. Flow cytometry 72 hpi quantified as fold change GFP MFI over WT (C). Each bar represents data for 3 independent sgRNAs (n = 3).

(D) Schematic of DNA damage recruitment of SMC5/6 complex proposed by (Räschle et al., 2015); Blue, KO increases unintegrated virus expression; red, no phenotype.

(E) Unintegrated virus reporter infection of clonal SLF1 and SLF2-KO cell lines, flow cytometry 48 hpi. Data from n = 3 quantified in (F). Error bars show standard deviation. ns, p > 0.05; ^∗∗∗p < 0.001.

See also Figures S2 and S4.