Search for novel factors affecting nuclear structure and function in Arabidopsis thaliana
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This thesis project searches for novel factors affecting nuclear structure and function in the plant model organism, Arabidopsis thaliana. The nuclear lamina is of critical importance to the cell as it has a multitude of roles. Its various functions include nuclear transport, involvement in signaling pathways, and chromatin organization. The structure of the nuclear lamina impacts its function, so an in depth understanding of the proteins composing the nuclear lamina is imperative. The well-studied animal nuclear lamina is composed of lamin, yet plants lack lamin orthologs. However, the plant nuclear lamina contains analogous proteins that are structurally and functionally similar. One of these proteins, CRWN4 is encoded by a member of a small family of genes required to maintain the structure of the plant nucleus. This thesis project utilizes a suppressor screen of the crwn4-2 mutation to find genes that could play a role in the structure and function of the plant nuclear lamina. The crwn4-2 allele is a missense mutation that reduces the abundance of the protein in the nucleus. Through this project, I identified a mutation that suppressed the crwn4-2 allele and restored normal protein levels in the nucleus. A series of genetic experiments were conducted to characterize the suppressor mutation and lay a foundation for its molecular identification. Through these genetic crosses, it was found that the suppressor allele is inherited in a semi-dominant manner and exhibits allele-specificity. Whole Genome Sequencing is currently being undertaken to locate the allele’s position within the genome and identify a candidate gene. The findings of this thesis provide evidence that genetic suppressor screens can be used to identify novel genes and interactions affecting plant nuclear architecture.