HelD is a Global Transcription Factor Enhancing Gene Expression in Rapidly Growing Mycobacteria

Vankova, V.; Kumar, D.; Shoman, M.; Schwarz, M.; Modrak, M.; Jirat Matejckova, J.; Neva, S.; Havelkova, J.; Sikova, M.; Pospsiilova, D.; Halada, P.; Sanderova, H.; Holubova, J.; Dohal, M.; Prevorovsky, M.; Stanek, O.; Knejzlik, Z.; Dvorakova, V.; Hnilicova, J.

doi:10.1101/2024.11.27.625628

bioRxiv · 10.1101/2024.11.27.625628

HelD is a Global Transcription Factor Enhancing Gene Expression in Rapidly Growing Mycobacteria

Vankova, V.·Kumar, D.·Shoman, M.·Schwarz, M.·Modrak, M.·Jirat Matejckova, J.·Neva, S.·Havelkova, J.·Sikova, M.·Pospsiilova, D.·Halada, P.·Sanderova, H.·Holubova, J.·Dohal, M.·Prevorovsky, M.·Stanek, O.·Knejzlik, Z.·Dvorakova, V.·Hnilicova, J.

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Abstract

HelD protein, also named HelR (encoded by MSMEG_2174 in Mycobacterium smegmatis), interacts with mycobacterial RNA polymerase (RNAP) and affects rifampicin resistance in Mycobacterium abscessus. Here, we provide data on rifampicin resistance and helD presence in the genomes of other clinically relevant nontuberculous mycobacteria. We show that helD is primarily found in rapidly growing mycobacteria, such as M. smegmatis, where we detected HelD at a subset of promoters that can also associate with CarD and RbpA. Transcriptome analysis of a helD deletion strain using RNA-seq revealed that HelD enhances gene expression during exponential growth and decreases it in stationary phase, during which we observed reduced levels of CarD, RbpA, and GTP, the initiation nucleotide for the majority of M. smegmatis transcripts. We propose a model in which HelD releases abortive RNAP complexes and confirm that HelD dissociates RNAP from the promoter in vitro. HelD not only helps mycobacteria overcome rifampicin treatment but also supports efficient transcription during rapid growth, which indicates a dual role of this transcription regulator.

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Vankova, V., Kumar, D., Shoman, M., Schwarz, M., Modrak, M., Jirat Matejckova, J., Neva, S., Havelkova, J., Sikova, M., Pospsiilova, D., Halada, P., Sanderova, H., Holubova, J., Dohal, M., Prevorovsky, M., Stanek, O., Knejzlik, Z., Dvorakova, V., Hnilicova, J.. 2024-11-28. HelD is a Global Transcription Factor Enhancing Gene Expression in Rapidly Growing Mycobacteria. https://doi.org/10.1101/2024.11.27.625628

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