Detailed Information on Publication Record
2023
Characterization of a transitionally occupied state and thermal unfolding of domain 1.1 of σ A factor of RNA polymerase from Bacillus subtilis
TUŽINČIN, Dávid, Petr PADRTA, Hana ŠANDEROVÁ, Alžbeta RABATINOVÁ, Kateřina BENDOVÁ et. al.Basic information
Original name
Characterization of a transitionally occupied state and thermal unfolding of domain 1.1 of σ A factor of RNA polymerase from Bacillus subtilis
Authors
TUŽINČIN, Dávid (703 Slovakia, belonging to the institution), Petr PADRTA (203 Czech Republic, belonging to the institution), Hana ŠANDEROVÁ, Alžbeta RABATINOVÁ, Kateřina BENDOVÁ (203 Czech Republic, belonging to the institution), Libor KRÁSNÝ, Lukáš ŽÍDEK (203 Czech Republic, guarantor, belonging to the institution) and Pavel KADEŘÁVEK (203 Czech Republic, belonging to the institution)
Edition
Proteins: Structure, Function and Bioinformatics, Wiley, 2023, 0887-3585
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10608 Biochemistry and molecular biology
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 2.900 in 2022
RIV identification code
RIV/00216224:14740/23:00131160
Organization unit
Central European Institute of Technology
UT WoS
001019240900001
Keywords in English
σA factor; Bacillus subtilis; NMR; RNA polymerase; conformational exchange.
Tags
International impact, Reviewed
Změněno: 27/10/2024 15:25, Ing. Martina Blahová
Abstract
V originále
σ factors are essential parts of bacterial RNA polymerase (RNAP) as they allow to recognize promotor sequences and initiate transcription. Domain 1.1 of vegetative σ factors occupies the primary channel of RNAP and also prevents binding of the σ factor to promoter DNA alone. Here, we show that domain 1.1 of Bacillus subtilis σ A exists in more structurally distinct variants in dynamic equilibrium. The major conformation at room temperature is represented by a previously reported well-folded structure solved by nuclear magnetic resonance (NMR), but 4% of the protein molecules are present in a less thermodynamically favorable state. We show that this population increases with temperature and we predict its significant elevation at higher but still biologically relevant temperatures. We characterized the minor state of the domain 1.1 using specialized methods of NMR. We found that, in contrast to the major state, the detected minor state is partially unfolded. Its propensity to form secondary structure elements is especially decreased for the first and third α helices, while the second α helix and β strand close to the C-terminus are more stable. We also analyzed thermal unfolding of the domain 1.1 and performed functional experiments with full length σ A and its shortened version lacking domain 1.1 ( σ A _ Δ 1.1 ). The results revealed that while full length σ A increases transcription activity of RNAP with increasing temperature, transcription with σ A _ Δ 1.1 remains constant. In summary, this study reveals conformational dynamics of domain 1.1 and provides a basis for studies of its interaction with RNAP and effects on transcription regulation.
Links
| EF18_070/0009846, research and development project |
| ||
| GA22-12023S, research and development project |
| ||
| GJ18-04197Y, research and development project |
| ||
| LX22NPO5103, research and development project |
| ||
| MUNI/A/1413/2022, interní kód MU |
| ||
| 90127, large research infrastructures |
|
