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Understanding the Regulation of Chromatin by Phase Separation

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dc.contributor.advisor Köhler, Alwin
dc.contributor.author SAHA, APURVA
dc.date.accessioned 2022-12-20T03:46:36Z
dc.date.available 2022-12-20T03:46:36Z
dc.date.issued 2022-12
dc.identifier.citation 53 en_US
dc.identifier.uri http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/7525
dc.description.abstract Chromatin is a highly dynamic DNA-protein complex that contributes to various essential processes in eukaryotes, including packaging of genomes. The fundamental unit of chromatin, the nucleosome core particle, comprises of the complex formed by DNA wrapped around the histone octamer (two copies each of H2A, H2B, H3, H4). Modifications of either the histones or DNA can regulate structure and function of chromatin, along with other downstream processes. Among them, the monoubiquitination of H2B modulate crucial pathways in various organisms. In yeast, Lge1 is a critical protein for monoubiquitination of the 123rd lysine of H2B and undergoes liquid-liquid phase separation. This physical property of Lge1 can concentrate the substrate chromatin and the ubiquitination machinery comprising of Rad6 (E2 conjugating enzyme) and Bre1 (E3 ligase) to give rise to condensed reaction chambers in vitro and is critical for maintaining optimal levels of ubiquitinated H2B in vivo. To elucidate the molecular mechanism of this process, single-molecule studies of reconstituted nucleosomal arrays, which have been used extensively to study chromatin and its interaction with different factors, can be done. We have optimized the reconstitution of nucleosome arrays using biotinylated λ phage DNA to generate a template to analyze how Lge1-Bre1 condensates encounter DNA that is in the form of chromatin. en_US
dc.description.sponsorship Medical University of Vienna, University of Vienna, KVPY en_US
dc.language.iso en en_US
dc.subject chromatin en_US
dc.subject curtain assays en_US
dc.subject reconstitution en_US
dc.subject single molecule imaging en_US
dc.title Understanding the Regulation of Chromatin by Phase Separation en_US
dc.type Thesis en_US
dc.description.embargo no embargo en_US
dc.type.degree BS-MS en_US
dc.contributor.department Dept. of Biology en_US
dc.contributor.registration 20171084 en_US


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  • MS THESES [1705]
    Thesis submitted to IISER Pune in partial fulfilment of the requirements for the BS-MS Dual Degree Programme/MSc. Programme/MS-Exit Programme

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