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Updated: Jul 17, 2026

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis
Published on: April 5, 2017
METTL14/YTHDF1 axis restores ULK1-mediated autophagic flux to suppress Mycobacterium tuberculosis survival
Mingjian Gao1, Shipeng Cao2, Huimin Liu3
1Department of Infectious Diseases, The Affiliated Changsha Central Hospital, Hengyang Medical School, University of South China, No.161 South Shaoshan Road, Yuhua District, Changsha, Hunan Province 410004, China.
Background:
Tuberculosis (TB) continues to be a major global infectious disease, and the ability of Mycobacterium tuberculosis (Mtb) to persist within macrophages is a major barrier to eradication. Autophagy is a critical host defense mechanism, yet the epitranscriptomic regulation of this process is poorly understood.
Methods:
PBMCs isolated from individuals with TB and THP-1 macrophages infected with Mtb H37Ra were analyzed for METTL14 expression. Functional modulation experiments were conducted to investigate autophagy, m6A modification, and bacterial survival. Techniques included qPCR, Western blot, immunofluorescence, Me-RIP, RIP, dual-luciferase reporter assays, RNA stability analysis, and CFU enumeration.
Results:
METTL14 level was significantly decreased in PBMCs of TB patients and in Mtb-infected macrophages. Overexpression of METTL14 restored autophagic flux, as evidenced by increased LC3 puncta, elevated Beclin-1 and ULK1 levels, and enhanced p62 degradation. Mechanistically, METTL14 mediated m6A modification of ULK1 mRNA, which was recognized by YTHDF1 to stabilize the transcript. Silencing ULK1 abrogated METTL14-induced autophagic flux, while pharmacological inhibition of autophagy reversed the suppression of intracellular Mtb survival by METTL14.
Conclusion:
These findings identify METTL14 as a key epitranscriptomic regulator of autophagic flux in macrophages during Mtb infection. By promoting m6A modification and stabilization of ULK1 mRNA, METTL14 restores autophagic flux and restricts bacterial persistence, highlighting a potential therapeutic target for TB.
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