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Roles of the Nucleosome Acidic Patch in Regulating Histone Modifications and Transcription

Cucinotta, Christine E. (2017) Roles of the Nucleosome Acidic Patch in Regulating Histone Modifications and Transcription. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

Proper transcription elongation is critical for all eukaryotes. One regulatory mechanism cells employ is modification of nucleosomes during transcription. H2B K123 monoubiquitylation (H2Bub) is a key histone posttranslational modification that correlates with transcription elongation, promotes downstream histone marks, and regulates chromatin architecture. The E2 and E3 enzymes, Rad6 and Bre1, catalyze H2Bub in collaboration with the Paf1 complex member Rtf1. Additionally, H2Bub and the histone chaperone complex FACT appear to be interdependent. While it is known that these factors promote and catalyze the modification, how these proteins interface with the nucleosome to promote transcription is still unclear. However, reports show that the nucleosome acidic patch is an important regulatory region that binds many different factors. This dissertation describes roles for the nucleosome acidic patch regulates the H2Bub modification cascade and transcription elongation efficiency in Saccharomyces cerevisiae. As the acidic patch is a hub for chromatin-binding proteins, I hypothesize that transcription elongation factors interface with the acidic patch to properly regulate gene expression. To determine whether the acidic patch may function directly in promoting H2Bub, I measured H2Bub levels in a minimal in vitro assay, and found that the acidic patch is required for proper H2Bub. To identify factors that bind to the nucleosome acidic patch in vivo, I implemented a proteomics approach that utilized site-specific crosslinking with which I uncovered an interaction between transcription elongation factors and the nucleosome acidic patch. These data, and that of others, show that the acidic patch can dynamically interact with chromatin-binding proteins to control gene expression.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Cucinotta, Christine E.cec71@pitt.eduCEC710000-0002-9644-3126
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairArndt, Karenarndt@pitt.eduarndt
Committee MemberVanDemark, Andrewandyv@pitt.eduandyv
Committee MemberRebeiz, Markrebeiz@pitt.edurebeiz
Committee MemberSchwacha, Anthonyschwacha@pitt.eduschwacha
Committee MemberMcManus, Charles J.mcmanus@andrew.cmu.edu
Date: 23 September 2017
Date Type: Publication
Defense Date: 12 July 2017
Approval Date: 23 September 2017
Submission Date: 2 August 2017
Access Restriction: 5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
Number of Pages: 215
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: chromatin, transcription, histone modifications, gene expression
Date Deposited: 24 Sep 2017 00:07
Last Modified: 24 Sep 2017 00:07
URI: http://d-scholarship.pitt.edu/id/eprint/32954

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