DNA binding ability of the Nse1-Nse3-Nse4 sub-complex of the
SMC5/6 complex is essential in fission yeast

a 2014

DNA binding ability of the Nse1-Nse3-Nse4 sub-complex of the SMC5/6 complex is essential in fission yeast

PALEČEK, Jan; Kateřina BEDNÁŘOVÁ; Hana SKOUPILOVÁ; Lenka JURČIŠINOVÁ; Lucie VONDROVÁ et al.

Základní údaje

Originální název

DNA binding ability of the Nse1-Nse3-Nse4 sub-complex of the SMC5/6 complex is essential in fission yeast

Autoři

Vydání

Yeast Chromosome Structure, Replication and Segregation, 2014

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10600 1.6 Biological sciences

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

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

Příznaky

Mezinárodní význam

Změněno: 20. 7. 2014 12:09, doc. Mgr. Jan Paleček, Dr. rer. nat.

Anotace

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. 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.

Návaznosti

GA13-00774S, projekt VaV

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

Investor: Grantová agentura ČR, Úloha proteinu Nse, podjednotek komplexu SMC5/6, v procesech stabilizujících pozastavené replikacní vidlice

Citovat

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 Yeast Chromosome Structure, Replication and Segregation. 2014.

@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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