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Silencing Motifs in the Clr2 Protein from Fission Yeast, Schizosaccharomyces pombe
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology. Uppsala University, Science for Life Laboratory, SciLifeLab.
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2014 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 9, no 1, e86948- p.Article in journal (Refereed) Published
Abstract [en]

The fission yeast, Schizosaccharomyces pombe, is a well-established model for heterochromatin formation, but the exact sequence of events for initiation remains to be elucidated. The essential factors involved include RNA transcribed from repeated sequences together with the methyltransferase Clr4. In addition, histone deacetylases, like Clr3, found in the SHREC complex are also necessary for transcriptional silencing. Clr2 is another crucial factor required for heterochromatin formation found in the SHREC complex. The function of Clr2 has been difficult to establish due to the lack of conserved domains or homology to proteins of known molecular function. Using a bioinformatics approach, three conserved motifs in Clr2 were identified, which contained amino acids important for transcriptional repression. Analysis of clr2 mutant strains revealed a major role for Clr2 in mating-type and rDNA silencing, and weaker effects on centromeric silencing. The effect on mating-type silencing showed variegation in several of the strains with mutated versions of Clr2 indicating an establishment or maintenance defect. Moreover, the critical amino acids in Clr2 were also necessary for transcriptional repression in a minimal system, by the tethering of Clr4 upstream of a reporter gene, inserted into the euchromatic part of the genome. Finally, in silico modeling suggested that the mutations in Clr2 cause disruption of secondary structures in the Clr2 protein. Identification of these critical amino acids in the protein provides a useful tool to explore the molecular mechanism behind the role of Clr2 in heterochromatin formation.

Place, publisher, year, edition, pages
2014. Vol. 9, no 1, e86948- p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-219965DOI: 10.1371/journal.pone.0086948ISI: 000330507300119OAI: oai:DiVA.org:uu-219965DiVA: diva2:704581
Available from: 2014-03-12 Created: 2014-03-09 Last updated: 2017-12-05Bibliographically approved
In thesis
1. Importance of the Clr2 protein in heterochromatin formation in the fission yeast Schizosaccharomyces pombe
Open this publication in new window or tab >>Importance of the Clr2 protein in heterochromatin formation in the fission yeast Schizosaccharomyces pombe
2017 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

Epigenetics is an area of biology that studies heritable changes in gene ex- pression without any change in the DNA sequence. The most studied epige- netic mechanisms are DNA methylation, RNA interference and histone mod- ifications. There are over 130 different modifications that can be attached to histones, and the most commonly studied are methylation, acetylation, phos- phorylation, sumoylation and ubiquitination. The modifications, spread out through the genome, form the histone code, which recruits transcription fac- tors and modifies the accessibility of the DNA, which results in either active or silenced transcription. The silenced form of chromatin is known as heter- ochromatin and is usually found in regions of the chromosome that need to be highly regulated. To study epigenetics, the model organism Schizosac- charomyces pombe is used widely used. S. pombe is a single cell, rod shaped, fission yeast. The simplicity of S. pombe and its similarities to high- er eukaryotes makes it a good model organism for studying epigenetics. We find that, when mutating evolutionary conserved amino acids in the Clr2 protein, which is involved in heterochromatin formation in S. pombe, there is a change in silencing in different heterochromatic regions. When constructs of Clr2 with the BAH domain deleted are overexpressed, there is an increase in silencing in the central core centromere of chromosome II of S. pombe. 

Place, publisher, year, edition, pages
Uppsala: Uppsala universitet, 2017. 73 p.
Keyword
Clr2, Yeast, Heterochromatin, Fission yeast, Schizosaccharomyces pombe
National Category
Medical and Health Sciences
Research subject
Biochemistry
Identifiers
urn:nbn:se:uu:diva-328296 (URN)
Presentation
2017-09-04, E10:2309, BMC, Husargatan 3, Uppsala, 10:15 (English)
Supervisors
Available from: 2017-08-21 Created: 2017-08-21 Last updated: 2017-09-15Bibliographically approved

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Steinhauf, DanielRodriguez, AlejandroMaksimov, VladimirBjerling, Pernilla

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