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Published on: July 19, 2021
Monitoring the mitochondrial localization of mycobacterial proteins
Krishnaveni Mohareer1, Jayashankar Medikonda1, Sriram Yandrapally1
1Department of Biochemistry, School of Life Sciences, University of Hyderabad, India.
Abstract:
Mitochondrion apart from being the energy hub of the cell, is also the center of various signaling pathways. During intracellular infection, either bacterial or viral, several pathogen proteins, metabolites, and possibly, lipids interact with the host mitochondria. These interactions allow the pathogens, such as Mtb, to reprogram the host mitochondrial functions, screwing the host immune responses and resulting in the persistence of the bacteria. Therefore, mitochondria offer a critical target organelle for various therapeutic interventions. This chapter deals with methods to demonstrate and establish the mitochondrial localization of Mtb proteins by confocal microscopy and mitochondrial enrichment. Transient transfection of mammalian constructs or infection Mycolicibacterium smegmatis infection could be used to overexpress the candidate Mtb protein in host cells, allowing the study of changes in the mitochondria's composition and function with regard to localization studies, mitochondrial DNA and RNA, proteomics, metabolomics, lipidomics, and bioenergetics.
Insights
Pathogen proteins can reprogram host mitochondria, aiding bacterial persistence. This study details methods to study Mycobacterium tuberculosis (Mtb) protein interactions with mitochondria for therapeutic insights.
Area of Science:
- Cell Biology
- Microbiology
- Immunology
Background:
- Mitochondria are central to cellular energy production and signaling.
- Intracellular pathogens like Mycobacterium tuberculosis (Mtb) interact with host mitochondria.
- These interactions reprogram mitochondrial function, impairing host immunity and promoting pathogen survival.
Purpose of the Study:
- To present methods for investigating Mtb protein localization within host mitochondria.
- To explore how Mtb proteins alter mitochondrial composition and function.
- To identify mitochondria as a therapeutic target for infectious diseases.
Main Methods:
- Confocal microscopy and mitochondrial enrichment techniques.
- Overexpression of candidate Mtb proteins in host cells via transfection or M. smegmatis infection.
- Analysis of mitochondrial DNA/RNA, proteomics, metabolomics, lipidomics, and bioenergetics.
Main Results:
- Established protocols for demonstrating Mtb protein mitochondrial localization.
- Enabled the study of pathogen-induced alterations in mitochondrial dynamics and function.
- Provided a framework for understanding host-pathogen interactions at the mitochondrial level.
Conclusions:
- Mitochondrial localization of Mtb proteins is crucial for bacterial persistence.
- Investigating these interactions offers avenues for novel therapeutic strategies.
- Mitochondria serve as a critical battleground in host-pathogen interactions.
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