Application of a designer histone platform to elucidate new roles for lysine methylation in chromatin regulation

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Lysine methylation is a signaling mechanism conserved from yeast to humans that is critical for many basic cellular processes. Moreover, these processes have been linked to many human diseases like developmental disorders and cancer. While many proteins related to lysine methylation signaling have been identified and characterized, many components of methyl-lysine signaling are poorly understood and require further mechanistic insight. In this work, we further to our understanding of methyl-lysine signaling by addressing technical and biological queries related to methyltransferase activity and identification and characterization of methyl-lysine binding proteins. In Chapter 2, we perform a systematic characterization of lysine methyltransferases on histone H3, a concentrated center for nuclear signaling, and put forth data highlighting the importance of nucleosomes when characterizing histone-modifying enzymes. In Chapter 3, we identify the PZP domain of AF10 to bind unmodified H3K27 and this binding event is regulated by modification of that residue. In cells, the binding interaction between H3 and AF10 regulates DOT1L and the deposition of H3K79 methylation, which manifests in leukemia cells sensitive to H3K79, suggesting a possible target for pharmaceutical intervention. H3K79 methylation is an important modification thought to be important for transcriptional elongation, however the mechanism linking these processes remains elusive. In Chapter 4, we suggest that the FACT complex may provide this link that can read H3K79 methylation signals. FACT is a histone chaperone complex important for relaxation chromatin ahead of RNA polymerase II to facilitate transcription elongation. Modest efficiency in remodeling H3K79me2 histones coupled with compensation of H3K79 depletion with increased FACT complex provide the strongest experimental links between H3K79 methylation and transcription elongation. Together, we contribute to the field of lysine methylation signaling by characterizing the biochemical and biological properties of writer and reader proteins with special consideration to how recombinant chromatin reagents may influence the results.


Type of resource text
Form electronic; electronic resource; remote
Extent 1 online resource.
Publication date 2016
Issuance monographic
Language English


Associated with Wilkinson, Alex W
Associated with Stanford University, Department of Biology.
Primary advisor Gozani, Or Pinchas
Thesis advisor Gozani, Or Pinchas
Thesis advisor Frydman, Judith
Thesis advisor Morrison, Ashby J
Thesis advisor Straight, Aaron, 1966-
Advisor Frydman, Judith
Advisor Morrison, Ashby J
Advisor Straight, Aaron, 1966-


Genre Theses

Bibliographic information

Statement of responsibility Alex W. Wilkinson.
Note Submitted to the Department of Biology.
Thesis Thesis (Ph.D.)--Stanford University, 2016.
Location electronic resource

Access conditions

© 2016 by Alex Wilkinson
This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).

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