The selective autophagy receptors Optineurin and p62 are both required for zebrafish host resistance to mycobacterial infection

PLoS Pathogens
Rui ZhangAnnemarie H Meijer

Abstract

Mycobacterial pathogens are the causative agents of chronic infectious diseases like tuberculosis and leprosy. Autophagy has recently emerged as an innate mechanism for defense against these intracellular pathogens. In vitro studies have shown that mycobacteria escaping from phagosomes into the cytosol are ubiquitinated and targeted by selective autophagy receptors. However, there is currently no in vivo evidence for the role of selective autophagy receptors in defense against mycobacteria, and the importance of autophagy in control of mycobacterial diseases remains controversial. Here we have used Mycobacterium marinum (Mm), which causes a tuberculosis-like disease in zebrafish, to investigate the function of two selective autophagy receptors, Optineurin (Optn) and SQSTM1 (p62), in host defense against a mycobacterial pathogen. To visualize the autophagy response to Mm in vivo, optn and p62 zebrafish mutant lines were generated in the background of a GFP-Lc3 autophagy reporter line. We found that loss-of-function mutation of optn or p62 reduces autophagic targeting of Mm, and increases susceptibility of the zebrafish host to Mm infection. Transient knockdown studies confirmed the requirement of both selective autophagy recepto...Continue Reading

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Methods Mentioned

BETA
transgenic
ubiquitination
PCR
electrophoresis
in vitro transcription
fluorescence microscopy
confocal microscopy

Software Mentioned

Image Lab
Fiji
Mega7
Clustal Omega
GraphPad Prism
Ensembl
GraphPad
ImageJ

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