Cited by 98Iron Metabolism and DisordersTuberculosis Research and EpidemiologyBacterial Genetics and Biotechnology

Iron-Dependent RNA-Binding Activity of Mycobacterium tuberculosis Aconitase

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Banerjee, Sharmistha, Nandyala, Ashok Kumar, Raviprasad, Podili, Ahmed, Niyaz, Hasnain, Seyed E.·2007·Journal of Bacteriology

Abstract

Cellular iron levels are closely monitored by iron regulatory and sensor proteins of Mycobacterium tuberculosis for survival inside macrophages. One such class of proteins systematically studied in eukaryotes and reported in a few prokaryotes are the iron-responsive proteins (IRPs). These IRPs bind to iron-responsive elements (IREs) present at untranslated regions (UTRs) of mRNAs and are responsible for posttranscriptional regulation of the expression of proteins involved in iron homeostasis. Amino acid sequence analysis of M. tuberculosis aconitase (Acn), a tricarboxylic acid (TCA) cycle enzyme, showed the presence of the conserved residues of the IRP class of proteins. We demonstrate that M. tuberculosis Acn is bifunctional. It is a monomeric protein that is enzymatically active in converting isocitrate to cis -aconitate at a broad pH range of 7 to 10 (optimum, pH 8). As evident from gel retardation assays, M. tuberculosis Acn also behaves like an IRP by binding to known mammalian IRE-like sequences and to predicted IRE-like sequences present at the 3′ UTR of thioredoxin ( trxC ) and the 5′ UTR of the iron-dependent repressor and activator ( ideR ) of M. tuberculosis. M. tuberculosis Acn when reactivated with Fe 2+ functions as a TCA cycle enzyme, but upon iron depletion by a specific iron chelator, it behaves like an IRP, binding to the selected IREs in vitro. Since iron is required for the Acn activity and inhibits the RNA-binding activity of Acn, the two activities of M. tuberculosis Acn are mutually exclusive. Our results demonstrate the bifunctional nature of M. tuberculosis Acn, pointing to its likely role in iron homeostasis.

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  1. [1]1. Ale ´n, C., and A. L. Sonenshein. 1999. Bacillus subtilis aconitase is an RNAbinding protein. Proc. Natl. Acad. Sci. USA 96: 10412-10417.

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Authors
Banerjee, S., Nandyala, A. K., Raviprasad, P., Ahmed, N., & Hasnain, S. E.
Affiliations
  • University of Hyderabad (IN)
  • Centre for DNA Fingerprinting and Diagnostics (IN)
  • Laboratory of Molecular and Cellular Biology, Centre for DNA Fingerprinting and Diagnostics, Hyderabad, 500076, India
  • Jawaharlal Nehru Centre for Advanced Scientific Research (IN)
Venue
Journal of Bacteriology
Published
2007-06
Volume
189
Issue
11
Pages
4046-4052
ISSN
  • 0021-9193 (ISSN-L)
  • 1098-5530
OpenAlex
W2123356497
Cited by
98
References
42
Citations by year
2025: 3 · 2024: 4 · 2023: 5 · 2022: 9 · 2021: 5 · 2020: 4 · 2019: 5 · 2018: 2 · 2017: 5 · 2016: 2 · 2015: 9 · 2014: 7 · 2013: 9 · 2012: 4
FWCI
3.64
Type
journal article
Language
en
Open access
Locations
  • Journal of Bacteriology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Council of Scientific and Industrial Research, India
  • Centre for DNA Fingerprinting and Diagnostics
Topics
  • Health Sciences › Medicine › Hematology › Iron Metabolism and Disorders
  • Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
Mycobacterium tuberculosis, aconitase, iron regulatory element, RNA-binding activity, iron homeostasis, post-transcriptional regulation, bifunctional protein, TCA cycle, thioredoxin, IdeR, iron depletion, iron chelation, macrophage survival
MeSH terms
Aconitate Hydratase, Bacterial Proteins, Chromatography, Gel, Hydrogen-Ion Concentration, Iron, Mycobacterium tuberculosis, Protein Binding, Recombinant Fusion Proteins, RNA, Substrate Specificity, Polymerase Chain Reaction, Open Reading Frames, RNA-Binding Proteins, Dimerization, Electrophoretic Mobility Shift Assay, Iron-Regulatory Proteins

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