Chromosomal association of the SMC5-6 complex is dependent on
interaction of its Nse1-Nse3-Nse4 subcomplex with DNA

a 2014

Chromosomal association of the SMC5-6 complex is dependent on interaction of its Nse1-Nse3-Nse4 subcomplex with DNA

ZÁBRADY, Kateřina; Marek ADAMUS; Hana SKOUPILOVÁ; Lenka JURČIŠINOVÁ; Chunyan LIAO et. al.

Basic information

Original name

Chromosomal association of the SMC5-6 complex is dependent on interaction of its Nse1-Nse3-Nse4 subcomplex with DNA

Name in Czech

Vazba komplexu SMC5-6 na chromatin je závislá na interakci jeho podkomplexu Nse1-Nse3-Nse4 s DNA

Authors

ZÁBRADY, Kateřina ; Marek ADAMUS; Hana SKOUPILOVÁ; Lenka JURČIŠINOVÁ; Chunyan LIAO; Alan R. LEHMANN and Jan PALEČEK

Edition

Annual Conference Brno 2014: Frontiers in Material and Life Sciences, 2014

Other information

Language

English

Type of outcome

Conference abstract

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

is not subject to a state or trade secret

Organization unit

Central European Institute of Technology

Keywords in English

SMC5/6 complex; NSE1/NSE3/NSE4 sub-complex; protein-DNA interaction; winged-helix motif; chromatin association

Changed: 12/3/2015 19:28, doc. Mgr. Jan Paleček, Dr. rer. nat.

Abstract

In the original language

SMC5-6 is a highly conserved protein complex related to cohesin and condensin complexes which are the key components of higher-order chromatin structures. The SMC5-6 complex is essential for proliferation in yeast and is involved in the homologous recombination-based DNA repair processes, including repair of DNA double strand breaks, restart of stalled replication forks etc. However, the precise mechanism of SMC5-6 function is not known. We will present the evidence for direct physical interaction of its part, Nse1-Nse3-Nse4 sub-complex, to DNA and its essential role for the function of the whole SMC5-6 complex. The Nse1-Nse3-Nse4 sub-complex is rich in winged-helix domain motifs and at least one of them form a putative DNA-binding cleft. The purified Nse1-Nse3-Nse4 sub-complexes shift different DNA substrates in electrophoretic mobility shift assays (EMSA) proving their ability to bind DNA in vitro. Mutations of the key basic residues within the putative DNA-binding cleft reduce in vitro binding to DNA. Introduction of these mutations into Schizosaccharomyces pombe genome results in cell death or hypersensitivity to hydroxyurea. The chromatin immuneprecipitation (ChIP) analysis of the DNA-binding mutant shows reduced association of SMC5-6 with chromatin. The above data and our genetic analysis indicate the essential role of the interaction between Nse1-Nse3-Nse4 and DNA for the loading and/or maintenance of the SMC5-6 complex on chromatin.

Links

CZ.1.05/1.1.00/02.0068, interní kód MU

Name: CEITEC - středoevropský technologický institut (Acronym: CEITEC)

Investor: Ministry of Education, Youth and Sports of the CR, CEITEC - Central European Institute of Technology, 1.1 European Centres of Excellence

GA13-00774S, research and development project

Name: Úloha proteinů Nse, podjednotek komplexu SMC5/6, v procesech stabilizujících pozastavené replikační vidlice

Investor: Czech Science Foundation

Cite

ZÁBRADY, Kateřina; Marek ADAMUS; Hana SKOUPILOVÁ; Lenka JURČIŠINOVÁ; Chunyan LIAO; Alan R. LEHMANN and Jan PALEČEK. Chromosomal association of the SMC5-6 complex is dependent on interaction of its Nse1-Nse3-Nse4 subcomplex with DNA. In Annual Conference Brno 2014: Frontiers in Material and Life Sciences. 2014.

@proceedings{1227920,
   author = {Zábrady, Kateřina and Adamus, Marek and Skoupilová, Hana and Jurčišinová, Lenka and Liao, Chunyan and Lehmann, Alan R. and Paleček, Jan},
   booktitle = {Annual Conference Brno 2014: Frontiers in Material and Life Sciences},
   keywords = {SMC5/6 complex; NSE1/NSE3/NSE4 sub-complex; protein-DNA interaction; winged-helix motif; chromatin association},
   language = {eng},
   title = {Chromosomal association of the SMC5-6 complex is dependent on interaction of its Nse1-Nse3-Nse4 subcomplex with DNA},
   year = {2014}
}

TY  - CONF
ID  - 1227920
AU  - Zábrady, Kateřina - Adamus, Marek - Skoupilová, Hana - Jurčišinová, Lenka - Liao, Chunyan - Lehmann, Alan R. - Paleček, Jan
PY  - 2014
TI  - Chromosomal association of the SMC5-6 complex is dependent on interaction of its Nse1-Nse3-Nse4 subcomplex with DNA
KW  - SMC5/6 complex
KW  - NSE1/NSE3/NSE4 sub-complex
KW  - protein-DNA interaction
KW  - winged-helix motif
KW  - chromatin association
N2  - SMC5-6 is a highly conserved protein complex related to cohesin and condensin complexes which are the key components of higher-order chromatin structures. The SMC5-6 complex is essential for proliferation in yeast and is involved in the homologous recombination-based DNA repair processes, including repair of DNA double strand breaks, restart of stalled replication forks etc. However, the precise mechanism of SMC5-6 function is not known. We will present the evidence for direct physical interaction of its part, Nse1-Nse3-Nse4 sub-complex, to DNA and its essential role for the function of the whole SMC5-6 complex. The Nse1-Nse3-Nse4 sub-complex is rich in winged-helix domain motifs and at least one of them form a putative DNA-binding cleft. The purified Nse1-Nse3-Nse4 sub-complexes shift different DNA substrates in electrophoretic mobility shift assays (EMSA) proving their ability to bind DNA in vitro. Mutations of the key basic residues within the putative DNA-binding cleft reduce in vitro binding to DNA. Introduction of these mutations into Schizosaccharomyces pombe genome results in cell death or hypersensitivity to hydroxyurea. The chromatin immuneprecipitation (ChIP) analysis of the DNA-binding mutant shows reduced association of SMC5-6 with chromatin. The above data and our genetic analysis indicate the essential role of the interaction between Nse1-Nse3-Nse4 and DNA for the loading and/or maintenance of the SMC5-6 complex on chromatin.
ER  -

ZÁBRADY, Kateřina; Marek ADAMUS; Hana SKOUPILOVÁ; Lenka JURČIŠINOVÁ; Chunyan LIAO; Alan R. LEHMANN and Jan PALEČEK. Chromosomal association of the SMC5-6 complex is dependent on interaction of its Nse1-Nse3-Nse4 subcomplex with DNA. In \textit{Annual Conference Brno 2014: Frontiers in Material and Life Sciences}. 2014.

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