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Journal of bacteriology
12 19, 2024;206 (12): e0006624.

MoaB2, a newly identified transcription factor, binds to �� in .

Barbora Brezovsk��, Subhash Narasimhan, Michaela ��ikov��, Hana ��anderov��, Tom���� Kova��, Nabajyoti Borah, Mahmoud Shoman, Debora Posp����ilov��, Viola Va��kov�� Hausnerov��, D��vid Tu��in��in, Martin ��ern��, Jan Kom��rek, Martina Janou��kov��, Milada Kambov��, Petr Halada, Alena K��enkov��, Martin Hub��lek, M��ria Trundov��, Jan Dohn��lek, Jarmila Hnilicov��, Luk���� ����dek, Libor Kr��sn��
Author Information
  1. Barbora Brezovsk��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia. ORCID
  2. Subhash Narasimhan: Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czechia.
  3. Michaela ��ikov��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  4. Hana ��anderov��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  5. Tom���� Kova��: Institute of Biotechnology of the Czech Academy of Sciences, Centre BIOCEV, Vestec, Czechia.
  6. Nabajyoti Borah: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  7. Mahmoud Shoman: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  8. Debora Posp����ilov��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  9. Viola Va��kov�� Hausnerov��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  10. D��vid Tu��in��in: Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czechia.
  11. Martin ��ern��: Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czechia.
  12. Jan Kom��rek: Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czechia.
  13. Martina Janou��kov��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  14. Milada Kambov��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia.
  15. Petr Halada: Institute of Microbiology of the Czech Academy of Sciences, Centre BIOCEV, Vestec, Czechia.
  16. Alena K��enkov��: Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Prague, Czechia.
  17. Martin Hub��lek: Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Prague, Czechia.
  18. M��ria Trundov��: Institute of Biotechnology of the Czech Academy of Sciences, Centre BIOCEV, Vestec, Czechia.
  19. Jan Dohn��lek: Institute of Biotechnology of the Czech Academy of Sciences, Centre BIOCEV, Vestec, Czechia.
  20. Jarmila Hnilicov��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia. ORCID
  21. Luk���� ����dek: Faculty of Science, National Centre for Biomolecular Research, Masaryk University, Brno, Czechia. ORCID
  22. Libor Kr��sn��: Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia. ORCID
PMID: 39499088 DOI: 10.1128/jb.00066-24

Abstract

In mycobacteria, �� is the primary sigma factor. This essential protein binds to RNA polymerase (RNAP) and mediates transcription initiation of housekeeping genes. Our knowledge about this factor in mycobacteria is limited. Here, we performed an unbiased search for interacting partners of ��. The search revealed a number of proteins; prominent among them was MoaB2. The ��-MoaB2 interaction was validated and characterized by several approaches, revealing that it likely does not require RNAP and is specific, as alternative �� factors (, closely related ��) do not interact with MoaB2. The structure of MoaB2 was solved by X-ray crystallography. By immunoprecipitation and nuclear magnetic resonance, the unique, unstructured N-terminal domain of �� was identified to play a role in the ��-MoaB2 interaction. Functional experiments then showed that MoaB2 inhibits ��-dependent (but not ��-dependent) transcription and may increase the stability of �� in the cell. We propose that MoaB2, by sequestering ��, has a potential to modulate gene expression. In summary, this study has uncovered a new binding partner of mycobacterial ��, paving the way for future investigation of this phenomenon.IMPORTANCEMycobacteria cause serious human diseases such as tuberculosis and leprosy. The mycobacterial transcription machinery is unique, containing transcription factors such as RbpA, CarD, and the RNA polymerase (RNAP) core-interacting small RNA Ms1. Here, we extend our knowledge of the mycobacterial transcription apparatus by identifying MoaB2 as an interacting partner of ��, the primary sigma factor, and characterize its effects on transcription and �� stability. This information expands our knowledge of interacting partners of subunits of mycobacterial RNAP, providing opportunities for future development of antimycobacterial compounds.

Keywords

MoaB2 RNA polymerase mycobacteria transcription ��A

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Grants

  1. 19-12956S/Czech Science Foundation
  2. 22-12023S/Czech Science Foundation
  3. 23-05622S/Czech Science Foundation
  4. LX22NPO5103/MEYS, Funded by the European Union-Next Generation EU
  5. RVO: 86652036/Czech Academy of Sciences

MeSH Term

Mycobacterium smegmatis
Sigma Factor
Bacterial Proteins
Gene Expression Regulation, Bacterial
Transcription Factors
Protein Binding
DNA-Directed RNA Polymerases
Crystallography, X-Ray
Transcription, Genetic

Chemicals

Sigma Factor
Bacterial Proteins
Transcription Factors
DNA-Directed RNA Polymerases
Journal Article Research Support, Non-U.S. Gov't
Article has an altmetric score of 23

Word Cloud

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