Transcription repressor protein ZBTB25 interacts with HDAC1 in macrophages infected with Mycobacterium tuberculosis, and its inhibition leads to autophagy and killing of the intracellular pathogen

Madhavan, A.; K B, A.; Raj Pushparajan, A.; M, B.; Ajay Kumar, R.

doi:10.1101/2020.06.22.164350

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bioRxiv · 10.1101/2020.06.22.164350

Transcription repressor protein ZBTB25 interacts with HDAC1 in macrophages infected with Mycobacterium tuberculosis, and its inhibition leads to autophagy and killing of the intracellular pathogen

Madhavan, A.·K B, A.·Raj Pushparajan, A.·M, B.·Ajay Kumar, R.

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Abstract

Downregulation of host gene expression is one of the key strategies adopted by intracellular pathogens such as Mycobacterium tuberculosis (MTB) for their survival and subsequent pathogenesis. In a previous study, we have shown that HDAC1 levels go up in macrophages infected with MTB and it hypoacetylates histone H3 at the promoter of IL-12B gene leading to its downregulation. Here we show that after infection with MTB, the levels of the phosphorylated form of HDAC1 increase significantly in macrophages. Employing immunoprecipitation and LC-MS/MS, we found transcriptional repressor protein ZBTB25 associates with HDAC1 silencing complex along with transcriptional corepressor Sin3a. By chromatin immunoprecipitation and PCR analyses, we found that phosphorylated HDAC1, Sin3a, and ZBTB25 are recruited to the promoter of IL-12B to downregulate its expression in infected macrophages. Knocking down of ZBTB25 enhanced release of IL-12p40 from infected macrophages. Interestingly, the treatment of infected macrophages with CI994 (inhibitor of HDAC1) or dithiopyridine (inhibitor of ZBTB25) promoted the colocalization of LC3 (microtubule-associated protein 1A/1B-light chain 3, a marker for autophagy) and MTB in autophagosomes. Induction of autophagy resulted in the killing of intracellular MTB. Enhanced phosphorylation of JAK2 and STAT4 was observed in macrophages upon CI994 and dithiopyridine treatment, and inhibition of JAK2/STAT4 negated the killing of intracellular mycobacteria suggesting a possible role of these proteins in the autophagy-mediated killing of intracellular MTB.

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Madhavan, A., K B, A., Raj Pushparajan, A., M, B., Ajay Kumar, R.. 2020-06-22. Transcription repressor protein ZBTB25 interacts with HDAC1 in macrophages infected with Mycobacterium tuberculosis, and its inhibition leads to autophagy and killing of the intracellular pathogen. https://doi.org/10.1101/2020.06.22.164350

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