Chromosomal association of the SMC5-6 complex is dependent on interaction of its Nse1-Nse3-Nse4 subcomplex with DNA
| Název česky | Vazba komplexu SMC5-6 na chromatin je závislá na interakci jeho podkomplexu Nse1-Nse3-Nse4 s DNA |
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| Autoři | |
| Rok publikování | 2014 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | 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. |
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