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@proceedings{1192751,
author = {Paleček, Jan and Bednářová, Kateřina and Skoupilová, Hana and Jurčišinová, Lenka and Vondrová, Lucie and Kolesár, Peter and Nováková, Markéta and Liao, Chunyan and Alt, Aaron and Oliver, Antony and QuangandDang, Hung and Murray, Johanne and Lehmann, Alan},
booktitle = {Yeast Chromosome Structure, Replication and Segregation},
keywords = {DNA binding, Nse1, Nse3, Nse4, SMC5-6 complex, DNA repair, chromatin structure},
language = {eng},
title = {DNA binding ability of the Nse1-Nse3-Nse4 sub-complex of the SMC5/6 complex is essential in fission yeast},
year = {2014}
}
TY - CONF
ID - 1192751
AU - Paleček, Jan - Bednářová, Kateřina - Skoupilová, Hana - Jurčišinová, Lenka - Vondrová, Lucie - Kolesár, Peter - Nováková, Markéta - Liao, Chunyan - Alt, Aaron - Oliver, Antony - Quang-Dang, Hung - Murray, Johanne - Lehmann, Alan
PY - 2014
TI - DNA binding ability of the Nse1-Nse3-Nse4 sub-complex of the SMC5/6 complex is essential in fission yeast
KW - DNA binding, Nse1, Nse3, Nse4, SMC5-6 complex, DNA repair, chromatin structure
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. The purified Nse1-Nse3-Nse4 sub-complexes shift different DNA substrates in electromobility 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 S. pombe genome results in cell death and/or hypersensitivity to hydroxyurea. The chromatin immunoprecipitaion (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 the yeast chromatin.
ER -
PALEČEK, Jan, Kateřina BEDNÁŘOVÁ, Hana SKOUPILOVÁ, Lenka JURČIŠINOVÁ, Lucie VONDROVÁ, Peter KOLESÁR, Markéta NOVÁKOVÁ, Chunyan LIAO, Aaron ALT, Antony OLIVER, Hung QUANG-DANG, Johanne MURRAY a Alan LEHMANN. DNA binding ability of the Nse1-Nse3-Nse4 sub-complex of the SMC5/6 complex is essential in fission yeast. In \textit{Yeast Chromosome Structure, Replication and Segregation}. 2014.
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