Domain 1.1 of σA factor of RNA polymerase from Bacillus subtilis beyond its major state conformation

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Domain 1.1 of σA factor of RNA polymerase from Bacillus subtilis beyond its major state conformation

Popis výsledku v původním jazyce

σ factors of RNA polymerase in bacteria are one of their key components which recognize promotor sequences and initiate transcription [1]. Domain 1.1 of σ A factors occupies the primary channel of RNA polymerase and prevents binding of the σ factor to promoter DNA [2,3]. Here we are presenting a study of domain 1.1 of σ A from the model organism B. subtilis and we are describing its dynamics at μs—ms timescale using detailed study of nuclear magnetic resonance together with its thermal unfolding characterized by optical and calorimetric methods. The results show that the domain 1.1 of σ A factor has natural ability to form a significantly less compact and more flexible state compared to the previously determined structure [4] at biologically relevant temperatures as documented by the secondary structure propensity (SSP) prediction based on the NMR data (Figure 1) and it is shown that even the hydrophobic core of the protein is affected by the transition of the state of the protein. The study is complemented with the functional characterization of the domain 1.1 in dependence on temperature. It clearly showed that the domain 1.1 is responsible for the response of the transcription process to an increase of the temperature supporting our hypothesis that the conformational plasticity of the domain 1.1 affects its ability to entry or exit the primary channel of RNA polymerase and it allows to increase the transcriptional output at higher temperatures. [1] Murakami K.S. and Darst S.A. (2003) Bacterial rna polymerases: the wholo story. Current opinion in structural biology, 13(1):31–39 [2] Bae B., Davis E., Brown D., Campbell E.A., Wigneshweraraj S., and Darst S.A. (2013). Phage t7 gp2 inhibition of escherichia coli rna polymerase involves misappropriation of σ 70 domain 1.1. Proceedings of the National Academy of Sciences, 110(49):19772–19777 [3] Murakami K.S. (2013) X-ray crystal structure of Escherichia coli RNA polymerase σ 70 holoenzyme. Journal of Biological Chemistry, 288(13):9126–9134

Název v anglickém jazyce

Domain 1.1 of σA factor of RNA polymerase from Bacillus subtilis beyond its major state conformation

Popis výsledku anglicky

σ factors of RNA polymerase in bacteria are one of their key components which recognize promotor sequences and initiate transcription [1]. Domain 1.1 of σ A factors occupies the primary channel of RNA polymerase and prevents binding of the σ factor to promoter DNA [2,3]. Here we are presenting a study of domain 1.1 of σ A from the model organism B. subtilis and we are describing its dynamics at μs—ms timescale using detailed study of nuclear magnetic resonance together with its thermal unfolding characterized by optical and calorimetric methods. The results show that the domain 1.1 of σ A factor has natural ability to form a significantly less compact and more flexible state compared to the previously determined structure [4] at biologically relevant temperatures as documented by the secondary structure propensity (SSP) prediction based on the NMR data (Figure 1) and it is shown that even the hydrophobic core of the protein is affected by the transition of the state of the protein. The study is complemented with the functional characterization of the domain 1.1 in dependence on temperature. It clearly showed that the domain 1.1 is responsible for the response of the transcription process to an increase of the temperature supporting our hypothesis that the conformational plasticity of the domain 1.1 affects its ability to entry or exit the primary channel of RNA polymerase and it allows to increase the transcriptional output at higher temperatures. [1] Murakami K.S. and Darst S.A. (2003) Bacterial rna polymerases: the wholo story. Current opinion in structural biology, 13(1):31–39 [2] Bae B., Davis E., Brown D., Campbell E.A., Wigneshweraraj S., and Darst S.A. (2013). Phage t7 gp2 inhibition of escherichia coli rna polymerase involves misappropriation of σ 70 domain 1.1. Proceedings of the National Academy of Sciences, 110(49):19772–19777 [3] Murakami K.S. (2013) X-ray crystal structure of Escherichia coli RNA polymerase σ 70 holoenzyme. Journal of Biological Chemistry, 288(13):9126–9134

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2023

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů