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Cited by 70Tuberculosis Research and EpidemiologyBacterial Genetics and BiotechnologyMycobacterium research and diagnosis

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Phosphorylation of Mycobacterial PcaA Inhibits Mycolic Acid Cyclopropanation

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Corrales, Rosa Milagros, Molle, Virginie, Leiba, Jade, Mourey, Lionel, de Chastellier, Chantal, Kremer, Laurent·2012·Journal of Biological Chemistry
DOI 10.1074/jbc.m112.373209PMC PMC3406704
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Abstract

Pathogenic mycobacteria survive within macrophages by residing in phagosomes, which they prevent from maturing and fusing with lysosomes. Although several bacterial components were seen to modulate phagosome processing, the molecular regulatory mechanisms taking part in this process remain elusive. We investigated whether the phagosome maturation block (PMB) could be modulated by signaling through Ser/Thr phosphorylation. Here, we demonstrated that mycolic acid cyclopropane synthase PcaA, but not MmaA2, was phosphorylated by mycobacterial Ser/Thr kinases at Thr-168 and Thr-183 both in vitro and in mycobacteria. Phosphorylation of PcaA was associated with a significant decrease in the methyltransferase activity, in agreement with the strategic structural localization of these two phosphoacceptors. Using a BCG ΔpcaA mutant, we showed that PcaA was required for intracellular survival and prevention of phagosome maturation in human monocyte-derived macrophages. The physiological relevance of PcaA phosphorylation was further assessed by generating PcaA phosphoablative (T168A/T183A) or phosphomimetic (T168D/T183D) mutants. In contrast to the wild-type and phosphoablative pcaA alleles, introduction of the phosphomimetic pcaA allele in the ΔpcaA mutant failed to restore the parental mycolic acid profile and cording morphotype. Importantly, the PcaA phosphomimetic strain, as the ΔpcaA mutant, exhibited reduced survival in human macrophages and was unable to prevent phagosome maturation. Our results add new insight into the importance of mycolic acid cyclopropane rings in the PMB and provide the first evidence of a Ser/Thr kinase-dependent mechanism for modulating mycolic acid composition and PMB.

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  • pcaA is transcriptionally upregulated in the active phase, and blocks phagosome-lysosome fusion and stimulation of the pro-inflammatory cytokine tumour necrosis factor in macrophage infections, which implies that cis cyclopropanations are important for early stages of infection.

    “pcaA is transcriptionally upregulated in the active phase60, and blocks phagosome–lysosome fusion96 and stimulation of the pro-​inflammatory cytokine tumour necrosis factor in macrophage infections97, which implies that cis cyclopropanations are impor- tant for early stages of infection.”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [96]

  • PknF inhibits PcaA by phosphorylating it, which may help to enforce dormancy in the chronic phase.

    “PknF inhibits PcaA by phosphorylating it96, which may help to enforce dormancy in the chronic phase94.”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [96]

  • PcaA is a cyclopropane synthase that adds the proximal cis-cyclopropanes of cis/cis-meromycolates.

    “PcaA is a cyclopropane synthase that adds the proximal cis-​cyclopropanes of cis/cis-​meromycolates96,97”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [96]

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Metadata

Authors
Corrales, R. M., Molle, V., Leiba, J., Mourey, L., de Chastellier, C., & Kremer, L.
Affiliations
Centre National de la Recherche Scientifique (FR)
Corresponding author
  • de Chastellier, Chantal
  • Kremer, Laurent
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2012-07
Volume
287
Issue
31
Pages
26187-26199
ISSN
0021-9258 (ISSN-L)
DOI
10.1074/jbc.m112.373209
PMC
PMC3406704
OpenAlex
W2019481782
Cited by
70
References
2
Citations by year
2026: 6 · 2025: 2 · 2023: 3 · 2022: 3 · 2021: 4 · 2020: 7 · 2019: 6 · 2018: 2 · 2017: 8 · 2016: 6 · 2015: 9 · 2014: 9 · 2013: 4 · 2012: 1
FWCI
5.10
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • HAL (submittedVersion)
  • HAL (submittedVersion, other-oa)
Funders
Agence Nationale de la Recherche (ANR-09-MIEN-0004)
Topics
  • Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
Keywords
mycobacteria, intracellular survival, macrophages, Ser/Thr kinase, phosphomimetic mutant
MeSH terms
Amino Acid Sequence, Bacterial Proteins, Cyclopropanes, Humans, Macrophages, Methyltransferases, Models, Molecular, Molecular Sequence Data, Mycobacterium bovis, Mycobacterium tuberculosis, Mycolic Acids, Phagosomes, Phosphorylation, Protein Processing, Post-Translational, Mutagenesis, Site-Directed, Conserved Sequence, Protein Serine-Threonine Kinases, Amino Acid Substitution, Microbial Viability, Host-Pathogen Interactions

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