Honors College Thesis

 

Mutation of Polycomb Repressive Complex 2 (PRC2) kmt6 to Probe in vivo Function in Fusarium Público Deposited

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https://ir.library.oregonstate.edu/concern/honors_college_theses/3n204141r

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  • Polycomb Repressive Complex 2 (PRC2) is a histone methyltransferase that initiates facultative heterochromatin formation by trimethylating histone H3 on lysine 27 (H3K27me3). Mutations within its three primary subunits, KMT6, EED, and SUZ12, result in inherited diseases and sporadic cancers. In the fungus Fusarium graminearum, deletion of the gene encoding the catalytic subunit, kmt6, results in derepression of ~20% of the genome as well as distinct phenotypes. A partial crystal structure for PRC2 from the fungus Chaetomium thermophilum has been published. Based on this, we generated KMT6 mutants, all tagged with GFP. Our hypothesis was that structure-function relationships proposed from the crystal structure will be supported by in vivo findings in our model fungus. After selectively mutating the primary sequence of kmt6 by in vitro site-directed mutagenesis, important catalytic functions and KMT6-EED-SUZ12 interactions were disturbed. As predicted, this led to many mutants exhibiting the Δkmt6 phenotype. However, most mutants exhibited both novel and diverse phenotypes, indicating distinct partial losses of function and/or differential silencing of the genome compared to both WT and Δkmt6.
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  • NSF (MCB1515998)
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  • 2017-07-27 to 2018-06-12

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