Cooperation between intrinsically disordered and ordered
regions of Spt6 regulates nucleosome and Pol II CTD binding,
and nucleosome assembly

J 2022

Cooperation between intrinsically disordered and ordered regions of Spt6 regulates nucleosome and Pol II CTD binding, and nucleosome assembly

KASILIAUSKAITE, Aiste, Karel KUBÍČEK, Tomáš KLUMPLER, Martina ZÁNOVÁ, David ZAPLETAL et. al.

Basic information

Original name

Cooperation between intrinsically disordered and ordered regions of Spt6 regulates nucleosome and Pol II CTD binding, and nucleosome assembly

Authors

KASILIAUSKAITE, Aiste (440 Lithuania, belonging to the institution), Karel KUBÍČEK (203 Czech Republic, belonging to the institution), Tomáš KLUMPLER (203 Czech Republic, belonging to the institution), Martina ZÁNOVÁ (703 Slovakia, belonging to the institution), David ZAPLETAL (203 Czech Republic, belonging to the institution), Eliska KOUTNA, Jiří NOVÁČEK (203 Czech Republic, belonging to the institution) and Richard ŠTEFL (203 Czech Republic, guarantor, belonging to the institution)

Edition

Nucleic acids research, Oxford, Oxford University Press, 2022, 0305-1048

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 14.900

RIV identification code

RIV/00216224:14740/22:00127507

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1093/nar/gkac451

UT WoS

000805243900001

Keywords in English

RNA polymerase II; transcription elongation factor; transcription elongation factor Spt6; unclassified drug

Abstract

V originále

Transcription elongation factor Spt6 associates with RNA polymerase II (Pol II) and acts as a histone chaperone, which promotes the reassembly of nucleosomes following the passage of Pol II. The precise mechanism of nucleosome reassembly mediated by Spt6 remains unclear. In this study, we used a hybrid approach combining cryo-electron microscopy and small-angle X-ray scattering to visualize the architecture of Spt6 from Saccharomyces cerevisiae. The reconstructed overall architecture of Spt6 reveals not only the core of Spt6, but also its flexible N- and C-termini, which are critical for Spt6’s function. We found that the acidic N-terminal region of Spt6 prevents the binding of Spt6 not only to the Pol II CTD and Pol II CTD-linker, but also to pre-formed intact nucleosomes and nucleosomal DNA. The N-terminal region of Spt6 self-associates with the tSH2 domain and the core of Spt6 and thus controls binding to Pol II and nucleosomes. Furthermore, we found that Spt6 promotes the assembly of nucleosomes in vitro. These data indicate that the cooperation between the intrinsically disordered and structured regions of Spt6 regulates nucleosome and Pol II CTD binding, and also nucleosome assembly.

Links

GA22-19896S, research and development project

Name: Strukturní podstata pro opětovné sestavení nukleosomu při přepisu genu zprostředkovaná proteinem Spt6

Investor: Czech Science Foundation, Structural basis for co-transcriptional nucleosome reassembly mediated by Spt6

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

649030, interní kód MU

Name: DECOR - Dynamic assembly and exchange of RNA polymerase II CTD factors (Acronym: DECOR)

Investor: European Union, DECOR, ERC (Excellent Science)

90127, large research infrastructures

Name: CIISB II

Citovat

KASILIAUSKAITE, Aiste, Karel KUBÍČEK, Tomáš KLUMPLER, Martina ZÁNOVÁ, David ZAPLETAL, Eliska KOUTNA, Jiří NOVÁČEK and Richard ŠTEFL. Cooperation between intrinsically disordered and ordered regions of Spt6 regulates nucleosome and Pol II CTD binding, and nucleosome assembly. Nucleic acids research. Oxford: Oxford University Press, 2022, vol. 50, No 10, p. 5961-5973. ISSN 0305-1048. Available from: https://dx.doi.org/10.1093/nar/gkac451.

@article{2240862,
   author = {Kasiliauskaite, Aiste and Kubíček, Karel and Klumpler, Tomáš and Zánová, Martina and Zapletal, David and Koutna, Eliska and Nováček, Jiří and Štefl, Richard},
   article_location = {Oxford},
   article_number = {10},
   doi = {http://dx.doi.org/10.1093/nar/gkac451},
   keywords = {RNA polymerase II; transcription elongation factor; transcription elongation factor Spt6; unclassified drug},
   language = {eng},
   issn = {0305-1048},
   journal = {Nucleic acids research},
   title = {Cooperation between intrinsically disordered and ordered regions of Spt6 regulates nucleosome and Pol II CTD binding, and nucleosome assembly},
   url = {https://academic.oup.com/nar/article/50/10/5961/6595227?login=true},
   volume = {50},
   year = {2022}
}

TY  - JOUR
ID  - 2240862
AU  - Kasiliauskaite, Aiste - Kubíček, Karel - Klumpler, Tomáš - Zánová, Martina - Zapletal, David - Koutna, Eliska - Nováček, Jiří - Štefl, Richard
PY  - 2022
TI  - Cooperation between intrinsically disordered and ordered regions of Spt6 regulates nucleosome and Pol II CTD binding, and nucleosome assembly
JF  - Nucleic acids research
VL  - 50
IS  - 10
SP  - 5961-5973
EP  - 5961-5973
PB  - Oxford University Press
SN  - 03051048
KW  - RNA polymerase II
KW  - transcription elongation factor
KW  - transcription elongation factor Spt6
KW  - unclassified drug
UR  - https://academic.oup.com/nar/article/50/10/5961/6595227?login=true
N2  - Transcription elongation factor Spt6 associates with RNA polymerase II (Pol II) and acts as a histone chaperone, which promotes the reassembly of nucleosomes following the passage of Pol II. The precise mechanism of nucleosome reassembly mediated by Spt6 remains unclear. In this study, we used a hybrid approach combining cryo-electron microscopy and small-angle X-ray scattering to visualize the architecture of Spt6 from Saccharomyces cerevisiae. The reconstructed overall architecture of Spt6 reveals not only the core of Spt6, but also its flexible N- and C-termini, which are critical for Spt6’s function. We found that the acidic N-terminal region of Spt6 prevents the binding of Spt6 not only to the Pol II CTD and Pol II CTD-linker, but also to pre-formed intact nucleosomes and nucleosomal DNA. The N-terminal region of Spt6 self-associates with the tSH2 domain and the core of Spt6 and thus controls binding to Pol II and nucleosomes. Furthermore, we found that Spt6 promotes the assembly of nucleosomes in vitro. These data indicate that the cooperation between the intrinsically disordered and structured regions of Spt6 regulates nucleosome and Pol II CTD binding, and also nucleosome assembly.
ER  -

KASILIAUSKAITE, Aiste, Karel KUBÍČEK, Tomáš KLUMPLER, Martina ZÁNOVÁ, David ZAPLETAL, Eliska KOUTNA, Jiří NOVÁČEK and Richard ŠTEFL. Cooperation between intrinsically disordered and ordered regions of Spt6 regulates nucleosome and Pol II CTD binding, and nucleosome assembly. \textit{Nucleic acids research}. Oxford: Oxford University Press, 2022, vol.~50, No~10, p.~5961-5973. ISSN~0305-1048. Available from: https://dx.doi.org/10.1093/nar/gkac451.

Detailed Information on Publication Record