Control of biotin biosynthesis in mycobacteria by a pyruvate carboxylase dependent metabolic signal Journal Article
| Authors: | Lazar, N.; Fay, A.; Nandakumar, M.; Boyle, K. E.; Xavier, J.; Rhee, K.; Glickman, M. S. |
| Article Title: | Control of biotin biosynthesis in mycobacteria by a pyruvate carboxylase dependent metabolic signal |
| Abstract: | Biotin is an essential cofactor utilized by all domains of life, but only synthesized by bacteria, fungi and plants, making biotin biosynthesis a target for antimicrobial development. To understand biotin biosynthesis in mycobacteria, we executed a genetic screen in Mycobacterium smegmatis for biotin auxotrophs and identified pyruvate carboxylase (Pyc) as required for biotin biosynthesis. The biotin auxotrophy of the pyc::tn strain is due to failure to transcriptionally induce late stage biotin biosynthetic genes in low biotin conditions. Loss of bioQ, the repressor of biotin biosynthesis, in the pyc::tn strain reverted biotin auxotrophy, as did reconstituting the last step of the pathway through heterologous expression of BioB and provision of its substrate DTB. The role of Pyc in biotin regulation required its catalytic activities and could be supported by M. tuberculosis Pyc. Quantitation of the kinetics of depletion of biotinylated proteins after biotin withdrawal revealed that Pyc is the most rapidly depleted biotinylated protein and metabolomics revealed a broad metabolic shift in wild type cells upon biotin withdrawal which was blunted in cell lacking Pyc. Our data indicate that mycobacterial cells monitor biotin sufficiency through a metabolic signal generated by dysfunction of a biotinylated protein of central metabolism. © 2017 John Wiley & Sons Ltd |
| Keywords: | controlled study; protein expression; unclassified drug; nonhuman; gene; protein depletion; transcription factor; enzyme activity; enzyme substrate; bacterial strain; biosynthesis; bacterial protein; mycobacterium tuberculosis; transcription regulation; enzyme analysis; catalysis; gene insertion; cell transport; genetic screening; transposon; biotinylation; biotin; bacterial gene; mycobacterium smegmatis; bacterial growth; metabolic activation; metabolomics; bacterial enzyme; bacterial cell; enzyme repression; biological monitoring; auxotrophy; enzyme defect; limit of quantitation; priority journal; article; biosynthetic gene; biotin derivative; desthiobiotin; protein bioa; protein biob; protein biod; protein biof; protein bioq; pyruvate carboxylase |
| Journal Title: | Molecular Microbiology |
| Volume: | 106 |
| Issue: | 6 |
| ISSN: | 0950-382X |
| Publisher: | Blackwell Publishing |
| Date Published: | 2017-12-01 |
| Start Page: | 1018 |
| End Page: | 1031 |
| Language: | English |
| DOI: | 10.1111/mmi.13865 |
| PROVIDER: | scopus |
| PUBMED: | 29052269 |
| PMCID: | PMC5916780 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 2 January 2018 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work