DNA binding ability of the Nse1-Nse3-Nse4 sub-complex is
critical for essential function of the SMC5-6 complex

a 2013

DNA binding ability of the Nse1-Nse3-Nse4 sub-complex is critical for essential function of the SMC5-6 complex

BEDNÁŘOVÁ, Kateřina, Lucie VONDROVÁ, Markéta NOVÁKOVÁ, Hana SKOUPILOVÁ, Chunian LIAO et. al.

Basic information

Original name

DNA binding ability of the Nse1-Nse3-Nse4 sub-complex is critical for essential function of the SMC5-6 complex

Authors

BEDNÁŘOVÁ, Kateřina, Lucie VONDROVÁ, Markéta NOVÁKOVÁ, Hana SKOUPILOVÁ, Chunian LIAO, Aron ALT, Antony OLIVER, Zdeněk KŘÍŽ, Johanne M. MURRAY, Alan LEHMANN and Jan PALEČEK

Edition

Chromatin, Replication and Chromosomal Stability 2013, 2013

Other information

Language

English

Type of outcome

Konferenční abstrakt

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

Organization unit

Central European Institute of Technology

Keywords in English

DNA binding, Nse1, Nse3, Nse4, SMC5-6 complex, DNA repair

Abstract

V originále

Smc5-6 is a highly conserved protein complex related to cohesin and condensin involved in the structural maintenance of chromosomes. 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 two of them form a putative DNA-binding cleft. Indeed, purified Nse1-Nse3-Nse4 sub-complexes shift different DNA substrates in electromobility shift assays proving their ability to bind DNA. Mutations of the key basic residues within the putative DNA-binding cleft result in significantly lower affinity to DNA compared to the wild-type sub-complexes suggesting that the proposed cleft mediates specific interactions with DNA substrates. Introduction of these mutations into S. pombe genome results in cell death and/or hypersensitivity to hydroxyurea. The lethal phenotype indicates that the interaction of the cleft with DNA is crucial for the essential function of the SMC5-6 complex.

Links

GA13-00774S, research and development project

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

Investor: Czech Science Foundation

Citovat

BEDNÁŘOVÁ, Kateřina, Lucie VONDROVÁ, Markéta NOVÁKOVÁ, Hana SKOUPILOVÁ, Chunian LIAO, Aron ALT, Antony OLIVER, Zdeněk KŘÍŽ, Johanne M. MURRAY, Alan LEHMANN and Jan PALEČEK. DNA binding ability of the Nse1-Nse3-Nse4 sub-complex is critical for essential function of the SMC5-6 complex. In Chromatin, Replication and Chromosomal Stability 2013. 2013.

@proceedings{1120476,
   author = {Bednářová, Kateřina and Vondrová, Lucie and Nováková, Markéta and Skoupilová, Hana and Liao, Chunian and Alt, Aron and Oliver, Antony and Kříž, Zdeněk and Murray, Johanne M. and Lehmann, Alan and Paleček, Jan},
   booktitle = {Chromatin, Replication and Chromosomal Stability 2013},
   keywords = {DNA binding, Nse1, Nse3, Nse4, SMC5-6 complex, DNA repair},
   language = {eng},
   title = {DNA binding ability of the Nse1-Nse3-Nse4 sub-complex is critical for essential function of the SMC5-6 complex},
   year = {2013}
}

TY  - CONF
ID  - 1120476
AU  - Bednářová, Kateřina - Vondrová, Lucie - Nováková, Markéta - Skoupilová, Hana - Liao, Chunian - Alt, Aron - Oliver, Antony - Kříž, Zdeněk - Murray, Johanne M. - Lehmann, Alan - Paleček, Jan
PY  - 2013
TI  - DNA binding ability of the Nse1-Nse3-Nse4 sub-complex is critical for essential function of the SMC5-6 complex
KW  - DNA binding, Nse1, Nse3, Nse4, SMC5-6 complex, DNA repair
N2  - Smc5-6 is a highly conserved protein complex related to cohesin and condensin involved in the structural maintenance of chromosomes. 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 two of them form a putative DNA-binding cleft. Indeed, purified Nse1-Nse3-Nse4 sub-complexes shift different DNA substrates in electromobility shift assays proving their ability to bind DNA. Mutations of the key basic residues within the putative DNA-binding cleft result in significantly lower affinity to DNA compared to the wild-type sub-complexes suggesting that the proposed cleft mediates specific interactions with DNA substrates. Introduction of these mutations into S. pombe genome results in cell death and/or hypersensitivity to hydroxyurea. The lethal phenotype indicates that the interaction of the cleft with DNA is crucial for the essential function of the SMC5-6 complex.
ER  -

BEDNÁŘOVÁ, Kateřina, Lucie VONDROVÁ, Markéta NOVÁKOVÁ, Hana SKOUPILOVÁ, Chunian LIAO, Aron ALT, Antony OLIVER, Zdeněk KŘÍŽ, Johanne M. MURRAY, Alan LEHMANN and Jan PALEČEK. DNA binding ability of the Nse1-Nse3-Nse4 sub-complex is critical for essential function of the SMC5-6 complex. In \textit{Chromatin, Replication and Chromosomal Stability 2013}. 2013.

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