Undergraduate Thesis Or Project
 

A Panel of Histone H3 Mutations to Investigate Centromere Maintenance and Gene Silencing

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  • Modifications of histone tail amino acid residues are necessary for many reactions involving chromatin. These processes can produce transcriptionally active, euchromatic, or inactive, heterochromatic, segments of DNA. The formation and maintenance of facultative heterochromatin is critical for the proper growth and development of many eukaryotes. Euchromatin can be changed into heterochromatin by post-translational modifications of core histone tail residues, for example, addition of methyl or acetyl functional groups to lysines or phosphate groups to serines and threonines. In two fungi, Neurospora crassa and Fusarium graminearum, the distinction between facultative and constitutive heterochromatin is maintained by methylation of histone H3 on lysine 27 (H3K27) and lysine 9 (H3K9), respectively. The H3 tail residues surrounding K9 (TARK9ST) and K27 (AARK27SA) are identical except for the underlined residues, threonine in the K9 region and alanine in the K27 region. Switching these regions may alter activity of the two methylating enzymes, KMT1 and KMT6, respectively. A large set of histone alleles with point mutations was created by an improved “Quick Change” method and standard cloning, and integrated into wildtype or mutant F. graminearum and N. crassa strains by targeted gene replacement of the normal hH3 gene. Verified mutants were screened for phenotypic changes and aberrant localization of the gene silencing machinery.
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  • National Science Foundation (MCB1515998) E.R. Jackman Friends and Alumni OSU STEM Leadership Academy, USDA Multicultural Scholars Program
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  • Existing Confidentiality Agreement
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  • 2017-11-08 to 2018-07-03
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