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.Základní údaje
Originální název
Characterization of a transitionally occupied state and thermal unfolding of domain 1.1 of σ A factor of RNA polymerase from Bacillus subtilis
Autoři
TUŽINČIN, Dávid (703 Slovensko, domácí), Petr PADRTA (203 Česká republika, domácí), Hana ŠANDEROVÁ, Alžbeta RABATINOVÁ, Kateřina BENDOVÁ (203 Česká republika, domácí), Libor KRÁSNÝ, Lukáš ŽÍDEK (203 Česká republika, garant, domácí) a Pavel KADEŘÁVEK (203 Česká republika, domácí)
Vydání
Proteins: Structure, Function and Bioinformatics, Wiley, 2023, 0887-3585
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
10608 Biochemistry and molecular biology
Stát vydavatele
Spojené státy
Utajení
není předmětem státního či obchodního tajemství
Odkazy
Impakt faktor
Impact factor: 2.900 v roce 2022
Kód RIV
RIV/00216224:14740/23:00131160
Organizační jednotka
Středoevropský technologický institut
UT WoS
001019240900001
Klíčová slova anglicky
σA factor; Bacillus subtilis; NMR; RNA polymerase; conformational exchange.
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 27. 10. 2024 15:25, Ing. Martina Blahová
Anotace
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.
Návaznosti
EF18_070/0009846, projekt VaV |
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GA22-12023S, projekt VaV |
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GJ18-04197Y, projekt VaV |
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LX22NPO5103, projekt VaV |
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MUNI/A/1413/2022, interní kód MU |
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90127, velká výzkumná infrastruktura |
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