New complexes of the bacterial transcription system

J. Srogoň1, T. Kovaž1, N. Borah2, T. Kouba3, P. Sudzinová2, H. Šanderová2, K. Hegrová2, M. Trundová1, J. Dušková1, T. Skálová1, K. Adámková1, L. Krásný2, J. Dohnálek1

1Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Vestec, Czech Republic

2Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic

3Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague,

Czech Republic

jirisrogon@gmail.com, tomas.koval@ibt.cas.cz

RNA polymerase, RNAP, is an enzyme complex responsible for synthesizing RNA molecules. RNAP copies RNA based on a DNA template during the process of transcription, a fundamental operation conserved from bacteria to humans.

Identifying specific interactions between RNAP and other proteins can lead to the discovery of new therapeutic targets for antibiotic development that are more effective and less prone to bacterial resistance. Additionally, understanding the mechanisms of resistance, which can arise from mutations altering interactions between RNA polymerase and other proteins, allows us to better address the problem of antibiotic resistance and develop new strategies to combat it [1, 2]. Finally, elucidating the role of RNAP and its protein complexes contributes to comprehensive understanding of bacterial biology.

Here, we discovered that a protein involved in protein translation that binds to RNAP, potentially linking the two processes in a not yet identified manner. Currently, we are characterizing the interactions between this protein and RNAP by biophysical and structural biology methods (cryo-EM, small-angle X-ray scattering, homologous modelling, etc.). In parallel, this interaction is being probed functionally by biochemical approaches.

1. Mosaei H, Harbottle J. Mechanisms of antibiotics inhibiting bacterial RNA polymerase. Biochem Soc Trans. (2019) 47, 339-350.

2. Sudzinová P, Šanderová H, Koval' T, Skálová T, Borah N, Hnilicová J, Kouba T, Dohnálek J, Krásný L. What the Hel: recent advances in understanding rifampicin resistance in bacteria. FEMS Microbiol Rev. (2023) 47, fuac051.

This work was supported by CSF (23-06295S), AS CR (86652036), MEYS (LM2023042), ERDF (CZ.02.1.01/0.0/0.0/18_046/0015974), and by support of Biocev CMS – core facilities Biophysical Methods, Structural Mass Spectrometry, Crystallization of Proteins and Nucleic Acids, Diffraction Techniques and of CEITEC, Masaryk University Cryo-electron microscopy and tomography core facility (CEMCOF), all of CIISB, part of Instruct-ERIC.