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Cited by 76Tuberculosis Research and EpidemiologyMycobacterium research and diagnosisBacterial Genetics and Biotechnology

Phosphorylation of KasB Regulates Virulence and Acid-Fastness in Mycobacterium tuberculosis

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Vilchèze, Catherine, Molle, Virginie, Carrère-Kremer, Séverine, Leiba, Jade, Mourey, Lionel, Shenai, Shubhada, Baronian, Grégory, Tufariello, Joann +9 more·2014·PLoS Pathogens
DOI 10.1371/journal.ppat.1004115PMC PMC4014462
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Abstract

Mycobacterium tuberculosis bacilli display two signature features: acid-fast staining and the capacity to induce long-term latent infections in humans. However, the mechanisms governing these two important processes remain largely unknown. Ser/Thr phosphorylation has recently emerged as an important regulatory mechanism allowing mycobacteria to adapt their cell wall structure/composition in response to their environment. Herein, we evaluated whether phosphorylation of KasB, a crucial mycolic acid biosynthetic enzyme, could modulate acid-fast staining and virulence. Tandem mass spectrometry and site-directed mutagenesis revealed that phosphorylation of KasB occurred at Thr334 and Thr336 both in vitro and in mycobacteria. Isogenic strains of M. tuberculosis with either a deletion of the kasB gene or a kasB_T334D/T336D allele, mimicking constitutive phosphorylation of KasB, were constructed by specialized linkage transduction. Biochemical and structural analyses comparing these mutants to the parental strain revealed that both mutant strains had mycolic acids that were shortened by 4-6 carbon atoms and lacked trans-cyclopropanation. Together, these results suggested that in M. tuberculosis, phosphorylation profoundly decreases the condensing activity of KasB. Structural/modeling analyses reveal that Thr334 and Thr336 are located in the vicinity of the catalytic triad, which indicates that phosphorylation of these amino acids would result in loss of enzyme activity. Importantly, the kasB_T334D/T336D phosphomimetic and deletion alleles, in contrast to the kasB_T334A/T336A phosphoablative allele, completely lost acid-fast staining. Moreover, assessing the virulence of these strains indicated that the KasB phosphomimetic mutant was attenuated in both immunodeficient and immunocompetent mice following aerosol infection. This attenuation was characterized by the absence of lung pathology. Overall, these results highlight for the first time the role of Ser/Thr kinase-dependent KasB phosphorylation in regulating the later stages of mycolic acid elongation, with important consequences in terms of acid-fast staining and pathogenicity.

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  • The FAS-II enzymes are all inhibited through phosphorylation by PknA, PknB and other STPKs.

    “The FAS-​II enzymes are all inhibited through phosphorylation by PknA, PknB and other STPKs89–93.”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [90]

    The Introduction (page not provided) says that most essential enzymes in the FASII central core are phosphorylated by STPKs and that phosphorylation inhibits their activity at least in vitro. The Results (page not provided) show that KasB can be phosphorylated by PknA, PknB and other kinases, but the paper does not establish that all FASII enzymes are inhibited by those kinases. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • PknH, PknD, PknL and PknF can also phosphorylate various FAS-II enzymes.

    “PknH, PknD, PknL and PknF can also phosphorylate various FAS-​II enzymes.”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [90]

    The Results section ('KasB is phosphorylated in vivo and in vitro on Thr334 and Thr336'; page unavailable) shows that PknH, PknD, PknL and PknF can phosphorylate KasB, a FAS-II enzyme. The Introduction names several other phosphorylated FAS-II enzymes, but does not identify these four kinases as phosphorylating those enzymes, so 'various FAS-II enzymes' is not fully supported. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • When KasB is inhibited by phosphorylation through PknF, M. tuberculosis loses its acid-fastness, has shorter meromycolate chains and a more permeable wall, and exhibits decreased uptake by macrophages and decreased growth and virulence in the mouse.

    “When KasB is inhibited by phosphorylation through PknF90, M. tuberculosis loses its acid-​fastness, has shorter mero- mycolate chains and a more permeable wall, and exhib- its decreased uptake by macrophages and decreased growth and virulence in the mouse90.”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [90]

    Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

    Experimental — accuracy not yet measured

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Authors
Vilchèze, C., Molle, V., Carrère-Kremer, S., Leiba, J., Mourey, L., Shenai, S., Baronian, G., Tufariello, J., Hartman, T., Veyron-Churlet, R., Trivelli, X., Tiwari, S., Weinrick, B., Alland, D., Guérardel, Y., Jacobs, W. R., & Kremer, L.
Affiliations
  • Albert Einstein College of Medicine (US)
  • Centre National de la Recherche Scientifique (FR)
  • Rutgers Health (US)
Corresponding author
Kremer, Laurent
ORCID
Venue
PLoS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2014-05-08
Volume
10
Issue
5
Pages
e1004115
ISSN
  • 1553-7366 (ISSN-L)
  • 1553-7374
DOI
10.1371/journal.ppat.1004115
PMC
PMC4014462
OpenAlex
W2025963602
Cited by
76
References
3
Citations by year
2026: 4 · 2025: 3 · 2024: 3 · 2023: 7 · 2022: 3 · 2021: 5 · 2020: 10 · 2019: 7 · 2018: 6 · 2017: 12 · 2016: 8 · 2015: 6 · 2014: 2
FWCI
6.56
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS Pathogens (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Rutgers University Community Repository (Rutgers University) (submittedVersion)
  • HAL (submittedVersion, cc-by)
  • HAL (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • LillOA (Université de Lille (University Of Lille)) (submittedVersion, cc-by)
Funders
  • Howard Hughes Medical Institute
  • Agence Nationale de la Recherche (ANR-09-MIEN-0004)
  • Inha University
  • Centre National de la Recherche Scientifique (IR-RMN-THC FR3050 CNRS, Fr3050, IR-RMN-THC Fr3050)
  • Université de Montpellier
  • National Institutes of Health (AI0-51519, AI26170)
  • National Institute of Allergy and Infectious Diseases (R01AI026170, AI-26170, P30AI051519, AI 051519, P01AI063537, R01AI098925, R37AI026170)
Topics
  • Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
  • Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
Mycobacterium tuberculosis, kasb, serine/threonine phosphorylation, mycolic acid biosynthesis, acid-fast staining, bacterial virulence, phosphomimetic mutant, catalytic triad, latent tuberculosis infection, aerosol infection, mouse model
MeSH terms
Animals, Bacterial Proteins, Cell Wall, Mice, Inbred C57BL, Microbiological Techniques, Models, Molecular, Mycobacterium tuberculosis, Mycolic Acids, Phosphorylation, Staining and Labeling, Tuberculosis, Virulence, 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase, Mice, SCID, Protein Serine-Threonine Kinases, Catalytic Domain, Lipid Metabolism, Mice

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