Mutations of CenH3 Domains and their Effect on Chromocenter Localization and Meiosis in Neurospora crassa Public

http://ir.library.oregonstate.edu/concern/honors_college_theses/hx11xh36r

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  • In recent years, non-coding DNA has received much attention within the scientific community. This attention has not only illuminated the mystery behind functionally important regions like the centromere, but has also brought to light additional important questions regarding centromere mechanisms, inheritance, and cellular recognition by the kinetochore, a protein complex that facilitates chromosome segregation. Although these questions have begun to be explored, centromeres still remain elusive, especially in the diverse and important group of filamentous fungi. The key finding that histones, the proteins that organize DNA, vary within centromere regions has led scientists to hypothesize that sequential variations hold the key to the observed altered function and kinetochore recognition. Compared to the normal histone H3, a centromere-specific H3 contains the most obvious sequence variations, especially in the N-terminal tail and the loop 1 region. Studies in humans have demonstrated that a two amino acid residue in the loop 1 region is important for retained centromere localization. In some filamentous fungi, the loop 1 addition is also present, prompting the scrutiny of this region for normal function. The N-terminus has also been shown to contain important element for normal meiotic function. To test these regions, loop 1 and N-terminal chimera were assembled in Neurospora crassa, a filamentous fungus. Point mutations were also introduced into the loop 1 region to check for functional importance. The N-terminal chimera demonstrated centromere localization while the loop 1 chimera was never successfully assembled. The loop 1 point mutants were successfully assembled and are currently in the process of being expressed in N. crassa.
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