Negative Regulation of Autophagy during Macrophage Infection by IMycobacterium bovis/I BCG via Protein Kinase C Activation

• Published in: International journal of molecular sciences Vol. 25; no. 6
• Main Authors: Maldonado-Bravo, Rafael, Villaseñor, Tomás, Pedraza-Escalona, Martha, Pérez-Martínez, Leonor, Hernández-Pando, Rogelio, Pedraza-Alva, Gustavo
• Format: Journal Article
• Language: English
• Published: MDPI AG 01.03.2024
• Subjects: BCG; BCG vaccines; Biological response modifiers; Cell death; Health aspects; Infection; Macrophages; Mass spectrometry; Protein kinases; Proteins; Tuberculosis; Mexico; Massachusetts
• DOI: 10.3390/ijms25063145
• ISSN: 1422-0067

Mycobacterium tuberculosis (Mtb) employs various strategies to manipulate the host’s cellular machinery, overriding critical molecular mechanisms such as phagosome-lysosome fusion, which are crucial for its destruction. The Protein Kinase C (PKC) signaling pathways play a key role in regulating phagocytosis. Recent research in Interferon-activated macrophages has unveiled that PKC phosphorylates Coronin-1, leading to a shift from phagocytosis to micropinocytosis, ultimately resulting in Mtb destruction. Therefore, this study aims to identify additional PKC targets that may facilitate Mycobacterium bovis (M. bovis) infection in macrophages. Protein extracts were obtained from THP-1 cells, both unstimulated and mycobacterial-stimulated, in the presence or absence of a general PKC inhibitor. We conducted an enrichment of phosphorylated peptides, followed by their identification through mass spectrometry (LC-MS/MS). Our analysis revealed 736 phosphorylated proteins, among which 153 exhibited alterations in their phosphorylation profiles in response to infection in a PKC-dependent manner. Among these 153 proteins, 55 are involved in various cellular processes, including endocytosis, vesicular traffic, autophagy, and programmed cell death. Importantly, our findings suggest that PKC may negatively regulate autophagy by phosphorylating proteins within the mTORC1 pathway (mTOR2/PKC/Raf-1/Tsc2/Raptor/Sequestosome-1) in response to M. bovis BCG infection, thereby promoting macrophage infection.