M. tuberculosis PknG manipulates host autophagy flux to promote pathogen intracellular survival
Open accessAbstract
The eukaryotic-type protein kinase G (PknG), one of the eleven eukaryotic type serine-threonine protein kinase (STPK) in Mycobacterium tuberculosis (Mtb), is involved in mycobacterial survival within macrophages, presumably by suppressing phagosome and autophagosome maturation, which makes PknG an attractive drug target. However, the exact mechanism by which PknG inhibits pathogen clearance during mycobacterial infection remains largely unknown. Here, we show that PknG promotes macroautophagy/autophagy induction but inhibits autophagosome maturation, causing an overall effect of blocked autophagy flux and enhanced pathogen intracellular survival. PknG prevents the activation of AKT (AKT serine/threonine kinase) via competitively binding to its pleckstrin homology (PH) domain, leading to autophagy induction. Remarkably, PknG could also inhibit autophagosome maturation to block autophagy flux via targeting host small GTPase RAB14. Specifically, PknG directly interacts with RAB14 to block RAB14-GTP hydrolysis. Furthermore, PknG phosphorylates TBC1D4/AS160 (TBC1 domain family member 4) to suppress its GTPase-activating protein (GAP) activity toward RAB14. In macrophages and in vivo, PknG promotes Mtb intracellular survival through blocking autophagy flux, which is dependent on RAB14. Taken together, our data unveil a dual-functional bacterial effector that tightly regulates host autophagy flux to benefit pathogen intracellular survival.Abbreviations: AKT: AKT serine/threonine kinase; ATG5: autophagy related 5; BMDMs: bone marrow-derived macrophages; DTT: dithiothreitol; FBS: fetal calf serum; GAP: GTPase-activating protein; MOI: multiplicity of infection; Mtb: Mycobacterium tuberculosis; MTOR: mechanistic target of rapamycin kinase; OADC: oleic acid-albumin-dextrose-catalase; PC, phosphatidylcholine; PH: pleckstrin homology; PI3K: phosphoinositide 3-kinase; PknG: protein kinase G; PtdIns(3,4,5)P3: phosphatidylinositol(3,4,5)-trisphosphate; SQSTM1: sequestosome 1; STPK: serine-threonine protein kinase; TB: tuberculosis; TBC1D4: TBC1 domain family member 4; TPR: tetratricopeptide repeat; ULK1: unc-51 like autophagy activating kinase 1; WT: wild-type
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PknG, in conjunction with the SapM phosphatase, blocks host phagosome maturation and modulates the host immune response, establishing PknG and SapM as bona fide virulence factors.
“This was subsequently validated in M. tuberculosis where PknG, in conjunction with the SapM phosphatase (86), blocks host phagosome maturation and modulates the host immune response (88, 96–98), establishing PknG and SapM as bona fide virulence factors.”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [97]
The section on PknG and RAB14 (p. 7) reports that PknG inhibits mycobacterial phagosome maturation and may thereby evade host immune clearance. It does not establish that PknG acts in conjunction with SapM or that SapM is a virulence factor; the citing sentence separately cites another source for SapM. (LLM, conf=0.87)
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- Authors
- Ge, P., Lei, Z., Yu, Y., Lu, Z., Qiang, L., Chai, Q., Zhang, Y., Zhao, D., Li, B., Pang, Y., Liu, C. H., & Wang, J.
- Affiliations
- Chinese Academy of Sciences (CN)
- CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Beijing, China
- Capital Medical University (CN)
- Corresponding author
- Liu, Cui Hua
- Wang, Jing
- ORCID
- Venue
- Autophagy
- Publisher
- Informa UK Limited
- Published
- 2021-07-07
- Volume
- 18
- Issue
- 3
- Pages
- 576-594
- ISSN
- 1554-8627 (ISSN-L)
- 1554-8635
- PMC
- PMC9037497
- OpenAlex
- W3197634257
- Cited by
- 132
- References
- 77
- Citations by year
- 2026: 36 · 2025: 38 · 2024: 28 · 2023: 18 · 2022: 10 · 2021: 2
- FWCI
- 4.59
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Autophagy (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Key Research and Development Program of China (2017YFD0500300)
- National Natural Science Foundation of China (31830003)
- Strategic Priority Research Program of the Chinese Academy of Sciences (XDB29020000)
- National Science and Technology Major Project (2018ZX10101004)
- Youth Innovation Promotion Association CAS (Y82R011CX3)
- the Open Project Program of the State Key Laboratory of Proteomics (SKLPO202003)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
- Keywords
- Mycobacterium tuberculosis, PknG, autophagic flux, autophagosome maturation, Rab14, Akt phosphorylation, pleckstrin homology domain, macrophages, intracellular survival, host-pathogen interaction, GTPase-activating protein
- MeSH terms
- Autophagy, Humans, Mycobacterium tuberculosis, Serine, Tuberculosis, Protein Serine-Threonine Kinases, Cyclic GMP-Dependent Protein Kinases, Phosphatidylinositol 3-Kinases, GTPase-Activating Proteins, rab GTP-Binding Proteins, Proto-Oncogene Proteins c-akt
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