1. Biochemistry and Chemical Biology
  2. Microbiology and Infectious Disease
Download icon

Mycobacterium tuberculosis induces decelerated bioenergetic metabolism in human macrophages

  1. Bridgette M Cumming
  2. Kelvin W Addicott
  3. John H Adamson
  4. Adrie JC Steyn  Is a corresponding author
  1. Africa Health Research Institute, South Africa
Tools and Resources
  • Cited 28
  • Views 3,744
  • Annotations
Cite this article as: eLife 2018;7:e39169 doi: 10.7554/eLife.39169

Abstract

How Mycobacterium tuberculosis (Mtb) rewires macrophage energy metabolism to facilitate survival is poorly characterized. Here, we used extracellular flux analysis to simultaneously measure the rates of glycolysis and respiration in real-time. Mtb infection induced a quiescent energy phenotype in human monocyte-derived macrophages and decelerated flux through glycolysis and the TCA cycle. In contrast, infection with the vaccine strain, M. bovis BCG, or dead Mtb induced glycolytic phenotypes with greater flux. Furthermore, Mtb reduced the mitochondrial dependency on glucose and increased the mitochondrial dependency on fatty acids, shifting this dependency from endogenous fatty acids in uninfected cells to exogenous fatty acids in infected macrophages. We demonstrate how quantifiable bioenergetic parameters of the host can be used to accurately measure and track disease, which will enable rapid quantifiable assessment of drug and vaccine efficacy. Our findings uncovered new paradigms for understanding the bioenergetic basis of host metabolic reprogramming by Mtb.

Article and author information

Author details

  1. Bridgette M Cumming

    Africa Health Research Institute, Durban, South Africa
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6977-765X
  2. Kelvin W Addicott

    Africa Health Research Institute, Durban, South Africa
    Competing interests
    The authors declare that no competing interests exist.
  3. John H Adamson

    Africa Health Research Institute, Durban, South Africa
    Competing interests
    The authors declare that no competing interests exist.
  4. Adrie JC Steyn

    Africa Health Research Institute, Durban, South Africa
    For correspondence
    adrie.steyn@ahri.org
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9177-8827

Funding

National Institutes of Health (R01AI111940)

  • Adrie JC Steyn

U.S. Department of Defense (PR121320)

  • Adrie JC Steyn

National Institutes of Health (R21127182)

  • Adrie JC Steyn

South African Medical Research Council

  • Adrie JC Steyn

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: Human monocytes were isolated from buffy coats bought from the South African National Blood Service with approval from SANBS Human Research Ethics Committee (Clearance Certificate No. 2016/02)

Reviewing Editor

  1. Bavesh D Kana, University of the Witwatersrand, South Africa

Publication history

  1. Received: June 13, 2018
  2. Accepted: November 15, 2018
  3. Accepted Manuscript published: November 16, 2018 (version 1)
  4. Version of Record published: December 7, 2018 (version 2)

Copyright

© 2018, Cumming et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,744
    Page views
  • 729
    Downloads
  • 28
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    Eric D Hoffer et al.
    Research Article Updated

    Modifications in the tRNA anticodon loop, adjacent to the three-nucleotide anticodon, influence translation fidelity by stabilizing the tRNA to allow for accurate reading of the mRNA genetic code. One example is the N1-methylguanosine modification at guanine nucleotide 37 (m1G37) located in the anticodon loop andimmediately adjacent to the anticodon nucleotides 34, 35, 36. The absence of m1G37 in tRNAPro causes +1 frameshifting on polynucleotide, slippery codons. Here, we report structures of the bacterial ribosome containing tRNAPro bound to either cognate or slippery codons to determine how the m1G37 modification prevents mRNA frameshifting. The structures reveal that certain codon–anticodon contexts and the lack of m1G37 destabilize interactions of tRNAPro with the P site of the ribosome, causing large conformational changes typically only seen during EF-G-mediated translocation of the mRNA-tRNA pairs. These studies provide molecular insights into how m1G37 stabilizes the interactions of tRNAPro with the ribosome in the context of a slippery mRNA codon.

    1. Biochemistry and Chemical Biology
    Eric M Jones et al.
    Tools and Resources

    In humans, the >800 G protein-coupled receptors (GPCRs) are responsible for transducing diverse chemical stimuli to alter cell state, and are the largest class of drug targets. Their myriad structural conformations and various modes of signaling make it challenging to understand their structure and function. Here we developed a platform to characterize large libraries of GPCR variants in human cell lines with a barcoded transcriptional reporter of G-protein signal transduction. We tested 7,800 of 7,828 possible single amino acid substitutions to the beta-2 adrenergic receptor (β2AR) at four concentrations of the agonist isoproterenol. We identified residues specifically important for β2AR signaling, mutations in the human population that are potentially loss of function, and residues that modulate basal activity. Using unsupervised learning, we resolve residues critical for signaling, including all major structural motifs and molecular interfaces. We also find a previously uncharacterized structural latch spanning the first two extracellular loops that is highly conserved across Class A GPCRs and is conformationally rigid in both the inactive and active states of the receptor. More broadly, by linking deep mutational scanning with engineered transcriptional reporters, we establish a generalizable method for exploring pharmacogenomics, structure and function across broad classes of drug receptors.