CoChat
Sign Up
Cited by 76Tuberculosis Research and EpidemiologyBacterial Genetics and BiotechnologyMicrobial Natural Products and Biosynthesis

The DOI matches its Crossref record; 4 of the 4 references it cites were found in a registry or an index.

Phosphorylation of Enoyl-Acyl Carrier Protein Reductase InhA Impacts Mycobacterial Growth and Survival

Open access
Khan, Shazia, Nagarajan, Sathya Narayanan, Parikh, Amit, Samantaray, Sharmishtha, Singh, Albel, Kumar, Devanand, Roy, Rajendra P., Bhatt, Apoorva, Nandicoori, Vinay Kumar·2010·Journal of Biological Chemistry
DOI 10.1074/jbc.m110.143131PMC PMC2988389
Open this paper in CoChat — free accountOpen PDF

Abstract

InhA, the primary target for the first line anti-tuberculosis drug isoniazid, is a key enzyme of the fatty-acid synthase II system involved in mycolic acid biosynthesis in Mycobacterium tuberculosis. In this study, we show that InhA is a substrate for mycobacterial serine/threonine protein kinases. Using a novel approach to validate phosphorylation of a substrate by multiple kinases in a surrogate host (Escherichia coli), we have demonstrated efficient phosphorylation of InhA by PknA, PknB, and PknH, and to a lower extent by PknF. Additionally, the sites targeted by PknA/PknB have been identified and shown to be predominantly located at the C terminus of InhA. Results demonstrate in vivo phosphorylation of InhA in mycobacteria and validate Thr-266 as one of the key sites of phosphorylation. Significantly, our studies reveal that the phosphorylation of InhA by kinases modulates its biochemical activity, with phosphorylation resulting in decreased enzymatic activity. Co-expression of kinase and InhA alters the growth dynamics of Mycobacterium smegmatis, suggesting that InhA phosphorylation in vivo is an important event in regulating its activity. An InhA-T266E mutant, which mimics constitutive phosphorylation, is unable to rescue an M. smegmatis conditional inhA gene replacement mutant, emphasizing the critical role of Thr-266 in mediating post-translational regulation of InhA activity. The involvement of various serine/threonine kinases in modulating the activity of a number of enzymes of the mycolic acid synthesis pathway, including InhA, accentuates the intricacies of mycobacterial signaling networks in parallel with the changing environment.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1074/jbc.m110.143131

Cites

4 of 4 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • 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 [92]

    The passage labeled Abstract says phosphorylation negatively affects KasA activity but enhances KasB activity. KasB is a FAS-II enzyme, so the paper contradicts the claim that all FAS-II enzymes are inhibited by phosphorylation. (LLM, conf=0.90)

    Checked by a reasoning model — may be wrong.

    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 [92]

    The Results report that PknH and, to a lesser extent, PknF phosphorylated the FAS-II enzyme InhA. Although PknD and PknL were included in the kinase screen, the paper does not report that either phosphorylated InhA or another FAS-II enzyme. The cited results therefore support only two of the four named kinases. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Through phosphorylation, PknH may substantially downregulate InhA activity in stasis, thus making M. tuberculosis less sensitive to isoniazid, as is observed in starvation.

    “through phosphorylation, PknH92 may substantially downregulate inha activity in stasis14, thus making M. tuberculosis less sensitive to isoniazid, as is observed in starvation17.”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [92]

    The Results report that PknH phosphorylates InhA, and the Discussion reports that phosphorylation decreases InhA’s enzymatic activity (Fig. 6). The paper does not establish that this downregulation occurs in stasis. The sentence separately cites other sources for PknH and for reduced isoniazid sensitivity during starvation. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 39

  1. Barry, 1998, Prog. Lipid Res., 37, 143, 10.1016/S0163-7827(98)00008-3 · DOI
  2. Takayama, 2005, Clin. Microbiol. Rev., 18, 81, 10.1128/CMR.18.1.81-101.2005 · DOI
  3. Bloch, 1977, Adv. Enzymol. Relat. Areas Mol. Biol., 45, 1
  4. Smith, 2003, Prog. Lipid Res., 42, 289, 10.1016/S0163-7827(02)00067-X · DOI
  5. Asselineau, 1950, Nature, 166, 782, 10.1038/166782a0 · DOI
  6. Kremer, 2001, J. Biol. Chem., 276, 27967, 10.1074/jbc.M103687200 · DOI
  7. Choi, 2000, J. Biol. Chem., 275, 28201, 10.1074/jbc.M003241200 · DOI
  8. Sacco, 2007, Proc. Natl. Acad. Sci. U.S.A., 104, 14628, 10.1073/pnas.0704132104 · DOI
  9. Marrakchi, 2000, Microbiology, 146, 289, 10.1099/00221287-146-2-289 · DOI
  10. Quémard, 1995, Biochemistry, 34, 8235, 10.1021/bi00026a004 · DOI
  11. Vilchèze, 2000, J. Bacteriol., 182, 4059, 10.1128/JB.182.14.4059-4067.2000 · DOI
  12. Bhatt, 2005, J. Bacteriol., 187, 7596, 10.1128/JB.187.22.7596-7606.2005 · DOI
  13. Cole, 1998, Nature, 393, 537, 10.1038/31159 · DOI · Paper page
  14. Av-Gay, 2000, Trends Microbiol., 8, 238, 10.1016/S0966-842X(00)01734-0 · DOI
  15. Chao, 2010, Biochim. Biophys. Acta, 1804, 620, 10.1016/j.bbapap.2009.09.008 · DOI
  16. Alber, 2009, Curr. Opin. Struct. Biol., 19, 650, 10.1016/j.sbi.2009.10.017 · DOI
  17. Cowley, 2004, Mol. Microbiol., 52, 1691, 10.1111/j.1365-2958.2004.04085.x · DOI
  18. Walburger, 2004, Science, 304, 1800, 10.1126/science.1099384 · DOI · Paper page
  19. Tiwari, 2009, J. Biol. Chem., 284, 27467, 10.1074/jbc.M109.036095 · DOI
  20. Papavinasasundaram, 2005, J. Bacteriol., 187, 5751, 10.1128/JB.187.16.5751-5760.2005 · DOI
  21. Deol, 2005, J. Bacteriol., 187, 3415, 10.1128/JB.187.10.3415-3420.2005 · DOI · Paper page
  22. Molle, 2006, J. Biol. Chem., 281, 30094, 10.1074/jbc.M601691200 · DOI · Paper page
  23. Bhatt, 2007, Mol. Microbiol., 64, 1442, 10.1111/j.1365-2958.2007.05761.x · DOI
  24. Veyron-Churlet, 2009, J. Biol. Chem., 284, 6414, 10.1074/jbc.M806537200 · DOI
  25. Kumar, 2009, J. Biol. Chem., 284, 11090, 10.1074/jbc.M808705200 · DOI
  26. Parikh, 2009, J. Mol. Biol., 386, 451, 10.1016/j.jmb.2008.12.031 · DOI
  27. Triccas, 1998, FEMS Microbiol. Lett., 167, 151, 10.1111/j.1574-6968.1998.tb13221.x · DOI
  28. Boyle, 1991, Methods Enzymol., 201, 110, 10.1016/0076-6879(91)01013-R · DOI
  29. Parish, 1998, Methods Mol. Biol., 101, 129
  30. Eblen, 2003, J. Biol. Chem., 278, 14926, 10.1074/jbc.M300485200 · DOI
  31. He, 2006, J. Med. Chem., 49, 6308, 10.1021/jm060715y · DOI
  32. Rozwarski, 1999, J. Biol. Chem., 274, 15582, 10.1074/jbc.274.22.15582 · DOI
  33. Molle, 2010, Mol. Microbiol., 75, 1064, 10.1111/j.1365-2958.2009.07041.x · DOI
  34. Canova, 2009, J. Bacteriol., 191, 2876, 10.1128/JB.01569-08 · DOI
  35. Greenstein, 2007, PLoS Pathog., 3, e49, 10.1371/journal.ppat.0030049 · DOI
  36. Sharma, 2006, FEBS J., 273, 2711, 10.1111/j.1742-4658.2006.05289.x · DOI · Paper page
  37. Cohen-Gonsaud, 2009, J. Biol. Chem., 284, 19290, 10.1074/jbc.M109.019653 · DOI
  38. Thakur, 2006, J. Biol. Chem., 281, 40107, 10.1074/jbc.M607216200 · DOI
  39. Snapper, 1990, Mol. Microbiol., 4, 1911, 10.1111/j.1365-2958.1990.tb02040.x · DOI · Paper page

Cited by

Sorted by citations

Metadata

Authors
Khan, S., Nagarajan, S. N., Parikh, A., Samantaray, S., Singh, A., Kumar, D., Roy, R. P., Bhatt, A., & Nandicoori, V. K.
Affiliations
  • National Institute of Immunology (IN)
  • University of Birmingham (GB)
  • University of Delhi (IN)
Corresponding author
Nandicoori, Vinay Kumar
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2010-11
Volume
285
Issue
48
Pages
37860-37871
ISSN
0021-9258 (ISSN-L)
DOI
10.1074/jbc.m110.143131
PMC
PMC2988389
OpenAlex
W2081977228
Cited by
76
References
4
Citations by year
2026: 1 · 2025: 3 · 2024: 4 · 2023: 3 · 2022: 3 · 2021: 2 · 2020: 6 · 2019: 6 · 2018: 5 · 2017: 10 · 2016: 5 · 2015: 6 · 2014: 10 · 2013: 3 · 2012: 5
FWCI
2.49
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)
  • University of Birmingham Research Portal (University of Birmingham) (submittedVersion)
  • Greater South Information System
Funders
Medical Research Council
Topics
  • Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › Microbial Natural Products and Biosynthesis
Keywords
InhA, FabI, protein phosphorylation, serine/threonine kinase, PknA, PknB, Mycobacterium tuberculosis, Mycobacterium smegmatis, mycolic acid biosynthesis, fatty acid synthase II, post-translational regulation, enzyme regulation
MeSH terms
Bacterial Proteins, Mycobacterium tuberculosis, Oxidoreductases, Phosphorylation, Protein Serine-Threonine Kinases, Microbial Viability

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review