Tale of tails; structural comparison of the RNA polymerase
δ-subunits

a 2024

Tale of tails; structural comparison of the RNA polymerase δ-subunits

ČERNÝ, Martin, Viktor BARTOŠÍK, Markéta PETRŮJOVÁ, Hana ŠANDEROVÁ, Libor KRÁSNÝ et. al.

Základní údaje

Originální název

Tale of tails; structural comparison of the RNA polymerase δ-subunits

Autoři

ČERNÝ, Martin, Viktor BARTOŠÍK, Markéta PETRŮJOVÁ, Hana ŠANDEROVÁ, Libor KRÁSNÝ a Lukáš ŽÍDEK

Vydání

XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB, 2024

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

Odkaz na Abstrakt

Organizační jednotka

Středoevropský technologický institut

ISSN

Změněno: 29. 4. 2024 14:00, Mgr. Martin Černý

Anotace

V originále

The δ-subunit of prokaryotic RNA polymerase is an integral part of the transcription machinery of many Gram-positive bacteria from the phylum Firmicutes. It was demonstrated that it plays critical role in the regulation of the stress responses, environmental adaptation, sporulation and virulence of several important pathogens, such as Staphylococcus aureus and Streptococcus spp. (1,2). It plays different roles in transcription; its part in the transcription termination and the RNA polymerase complex recycling was recently well described (3). On the other hand, the exact mechanism of function during the transcription initiation are still not fully understood. In the past, our group was able to solve the structure of δ-subunit from Bacillus subtilis. The ~20 kDa protein is comprised of a structured N-terminal domain (NTD) and a negatively charged and poorly conserved C-terminal domain (CTD), with an exception of the well-conserved lysine tract at the beginning of the CTD. We previously demonstrated, that the negatively charged CTD transiently interacts with the lysines, leading to a more compacted structure of the δ-subunit (4). However, some of the Firmicutes, such as S. aureus, are missing the lysine tract which is instead located on the RNA polymerase core. In this poster, we designed purification protocols to obtain pure samples of both proteins and their respective mutants (devoid or with lysine tract included) from B. subtilis and S. aureus. We then compared their structural properties by means of circular dichroism spectroscopy (CD), small-angle X-ray scattering (SAXS) and nuclear magnetic resonance spectroscopy (NMR) to probe the importance of the lysine tract for the global structure of δ-subunit.

Citovat

ČERNÝ, Martin, Viktor BARTOŠÍK, Markéta PETRŮJOVÁ, Hana ŠANDEROVÁ, Libor KRÁSNÝ a Lukáš ŽÍDEK. Tale of tails; structural comparison of the RNA polymerase δ-subunits. In XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB. 2024. ISSN 1211-5894.

@proceedings{2398057,
   author = {Černý, Martin and Bartošík, Viktor and Petrůjová, Markéta and Šanderová, Hana and Krásný, Libor and Žídek, Lukáš},
   booktitle = {XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB},
   language = {eng},
   title = {Tale of tails; structural comparison of the RNA polymerase δ-subunits},
   url = {https://www.xray.cz/setkani/abst2024/773.htm},
   year = {2024}
}

TY  - CONF
ID  - 2398057
AU  - Černý, Martin - Bartošík, Viktor - Petrůjová, Markéta - Šanderová, Hana - Krásný, Libor - Žídek, Lukáš
PY  - 2024
TI  - Tale of tails; structural comparison of the RNA polymerase δ-subunits
UR  - https://www.xray.cz/setkani/abst2024/773.htm
N2  - The δ-subunit of prokaryotic RNA polymerase is an integral part of the transcription machinery of many Gram-positive bacteria from the phylum Firmicutes. It was demonstrated that it plays critical role in the regulation of the stress responses, environmental adaptation, sporulation and virulence of several important pathogens, such as Staphylococcus aureus and Streptococcus spp. (1,2). It plays different roles in transcription; its part in the transcription termination and the RNA polymerase complex recycling was recently well described (3). On the other hand, the exact mechanism of function during the transcription initiation are still not fully understood. In the past, our group was able to solve the structure of δ-subunit from Bacillus subtilis. The ~20 kDa protein is comprised of a structured N-terminal domain (NTD) and a negatively charged and poorly conserved C-terminal domain (CTD), with an exception of the well-conserved lysine tract at the beginning of the CTD. We previously demonstrated, that the negatively charged CTD transiently interacts with the lysines, leading to a more compacted structure of the δ-subunit (4). However, some of the Firmicutes, such as S. aureus, are missing the lysine tract which is instead located on the RNA polymerase core. In this poster, we designed purification protocols to obtain pure samples of both proteins and their respective mutants (devoid or with lysine tract included) from B. subtilis and S. aureus. We then compared their structural properties by means of circular dichroism spectroscopy (CD), small-angle X-ray scattering (SAXS) and nuclear magnetic resonance spectroscopy (NMR) to probe the importance of the lysine tract for the global structure of δ-subunit.
ER  -

ČERNÝ, Martin, Viktor BARTOŠÍK, Markéta PETRŮJOVÁ, Hana ŠANDEROVÁ, Libor KRÁSNÝ a Lukáš ŽÍDEK. Tale of tails; structural comparison of the RNA polymerase δ-subunits. In \textit{XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB}. 2024. ISSN~1211-5894.

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