Phosphoproteomics reveals new insights into the role of PknG during the persistence of pathogenic mycobacteria in host macrophages

摘要: Abstract Pathogenic mycobacteria, such as Mycobacterium tuberculosis, modulate the host immune system to evade clearance and promote long-term persistence, resulting in disease progression or latent infection. Understanding mechanisms pathogenic mycobacteria use escape elimination by is critical better understanding molecular of mycobacterial Protein kinase G (PknG) has been shown play an important role avoiding macrophages through blocking phagosome-lysosome fusion; however, exact mechanism not completely understood. Here, further investigate PknG during early events macrophage infection, RAW 264.7 cell lines were infected with M. bovis BCG wild-type knock-out mutant strains. After proteolysis, phosphopeptides enriched via TiO2 columns subjected LC-MS/MS identify differentially phosphorylated peptides between macrophages. A total 1401 phosphosites on 914 unique proteins identified. Following phosphoproteome normalisation differential expression analysis, a 149 versus mutant. subset 95 was up-regulated presence PknG. Functional analysis our data revealed that activity reprograms normal function interfering cytoskeletal organisation, spliceosomal machinery, translational initiation, programmed death. Differentially this study serve foundation for validation substrate assignment. Importance Tuberculosis (TB) remains one leading causes death from infection worldwide, due ability tuberculosis (Mtb) survive replicate within host, establishing reservoirs live bacteria persistence recurrence disease. Mtb uses thus major goal TB field. thought disruption function, but underlying are well new phosphoproteomic reveals substantially extensive PknG-mediated post-translational control cellular processes. These novel findings therefore considerably increase knowledge pathogenicity, including specific pathways might be re-activatable host-directed therapy, thereby restoring eliminate Mtb.