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The SMC5/6 complex leads to a loss of H3K27ac on KSHV chromatin.

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posted on 01.08.2022, 17:48 authored by Chunyan Han, Dun Zhang, Chenwu Gui, Liang Huang, Sijia Chang, Lianghui Dong, Lei Bai, Shuwen Wu, Ke Lan

(a-c) Ectopic expression of SMC6 decreases the levels of H3K27ac on KSHV genome. (a) H3K27ac on viral RTA was measured with ChIP-qPCR assay. ChIP was performed among latently infected control cells and SMC6-overexpressing cells by using antibodies against total H3 or H3K27ac. The recruitment of H3 and H3K27ac on KSHV RTA were tested via qPCR. Data was calculated as the fold change in percentage of input DNA compared with control ChIP experiment. (b) H3K27ac on viral LANA was measured with ChIP-qPCR assay. (c) H3K27ac on viral TR was measured with ChIP-qPCR assay. (d-g) Knockdown of SMC6 increases the levels of H3K27ac on KSHV genome. (d) iSLK.RGB cells were transduced with lentivirus expressing shRNA against SMC6. The knockdown efficiency was determined by western blots. (e-g) H3K27ac on viral RTA, LANA and TR were measured with ChIP-qPCR assay. ChIP was performed among latently infected control cells and SMC6 knockdown cells by using antibodies against H3 or H3K27ac. The recruitment of H3 and H3K27ac on KSHV genome were tested via qPCR. Representative results from three biological replicates are presented. Error bars indicate SD. Data were analyzed with Student’s t tests (**p< 0.01, ***p< 0.001).

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