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Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae

The silent mating-type loci HML and HMR of Saccharomyces cerevisiae contain mating-type information that is permanently repressed. This silencing is mediated by flanking sequence elements, the E- and I-silencers. They contain combinations of binding sites for the proteins Rap1, Abf1 and Sum1 as well...

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Autores principales: Weber, Jan M., Ehrenhofer-Murray, Ann E.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3001064/
https://www.ncbi.nlm.nih.gov/pubmed/20699276
http://dx.doi.org/10.1093/nar/gkq689
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author Weber, Jan M.
Ehrenhofer-Murray, Ann E.
author_facet Weber, Jan M.
Ehrenhofer-Murray, Ann E.
author_sort Weber, Jan M.
collection PubMed
description The silent mating-type loci HML and HMR of Saccharomyces cerevisiae contain mating-type information that is permanently repressed. This silencing is mediated by flanking sequence elements, the E- and I-silencers. They contain combinations of binding sites for the proteins Rap1, Abf1 and Sum1 as well as for the origin recognition complex (ORC). Together, they recruit other silencing factors, foremost the repressive Sir2/Sir3/Sir4 complex, to establish heterochromatin-like structures at the HM loci. However, the HM silencers exhibit considerable functional redundancy, which has hampered the identification of further silencing factors. In this study, we constructed a synthetic HML-E silencer (HML-SS ΔI) that lacked this redundancy. It consisted solely of Rap1 and ORC-binding sites and the D2 element, a Sum1-binding site. All three elements were crucial for minimal HML silencing, and mutations in these elements led to a loss of Sir3 recruitment. The silencer was sensitive to a mutation in RAP1, rap1-12, but less sensitive to orc mutations or sum1Δ. Moreover, deletions of SIR1 and DOT1 lead to complete derepression of the HML-SS ΔI silencer. This fully functional, minimal HML-E silencer will therefore be useful to identify novel factors involved in HML silencing.
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spelling pubmed-30010642010-12-13 Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae Weber, Jan M. Ehrenhofer-Murray, Ann E. Nucleic Acids Res Gene Regulation, Chromatin and Epigenetics The silent mating-type loci HML and HMR of Saccharomyces cerevisiae contain mating-type information that is permanently repressed. This silencing is mediated by flanking sequence elements, the E- and I-silencers. They contain combinations of binding sites for the proteins Rap1, Abf1 and Sum1 as well as for the origin recognition complex (ORC). Together, they recruit other silencing factors, foremost the repressive Sir2/Sir3/Sir4 complex, to establish heterochromatin-like structures at the HM loci. However, the HM silencers exhibit considerable functional redundancy, which has hampered the identification of further silencing factors. In this study, we constructed a synthetic HML-E silencer (HML-SS ΔI) that lacked this redundancy. It consisted solely of Rap1 and ORC-binding sites and the D2 element, a Sum1-binding site. All three elements were crucial for minimal HML silencing, and mutations in these elements led to a loss of Sir3 recruitment. The silencer was sensitive to a mutation in RAP1, rap1-12, but less sensitive to orc mutations or sum1Δ. Moreover, deletions of SIR1 and DOT1 lead to complete derepression of the HML-SS ΔI silencer. This fully functional, minimal HML-E silencer will therefore be useful to identify novel factors involved in HML silencing. Oxford University Press 2010-12 2010-08-10 /pmc/articles/PMC3001064/ /pubmed/20699276 http://dx.doi.org/10.1093/nar/gkq689 Text en © The Author(s) 2010. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Gene Regulation, Chromatin and Epigenetics
Weber, Jan M.
Ehrenhofer-Murray, Ann E.
Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae
title Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae
title_full Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae
title_fullStr Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae
title_full_unstemmed Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae
title_short Design of a minimal silencer for the silent mating-type locus HML of Saccharomyces cerevisiae
title_sort design of a minimal silencer for the silent mating-type locus hml of saccharomyces cerevisiae
topic Gene Regulation, Chromatin and Epigenetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3001064/
https://www.ncbi.nlm.nih.gov/pubmed/20699276
http://dx.doi.org/10.1093/nar/gkq689
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