Interaction and Functional Analysis of Drosophila Mcm10
In eukaryotic cells, DNA replication and the subsequent packaging of DNA into specific chromatin states are essential processes for the transmission of genetic material and genomic stability which may be linked by interactions between DNA replication proteins and chromatin association proteins. The discovery of several proteins with roles in both processes is evidence for this. Mcm10 is one such protein that has shown evidence for multiple roles in DNA replication, heterochromatin formation, and chromosome condensation. The separation of the functions of Mcm10 are investigated in this study by the analysis of two mutant alleles. A hypomorphic allele of Mcm10 demonstrates that Mcm10 has a role in heterochromatic silencing, chromosome condensation, and DNA replication. The analysis of a C terminal truncation allele indicates a role in endoreplication mediated through an interaction with Mcm2. The C terminal however, does not seem to be involved in heterochromatic silencing or chromosome condensation. The second phase of this study involves the development of a novel yeast three-hybrid system. The yeast two-hybrid system is a useful tool for detecting interactions between two proteins and identifying novel protein interactions. However, one limitation to the system is that some two-protein interactions require a third protein to stabilize or facilitate the binding between the two. In this system, we present two novel vectors; pGBKTet and pHook (derived from pDela) that are Gatewaytm compatible and allow screening for the third protein. Due to its numerous protein interactions and its exceptional abundance in the eukaryotic cell with approximately 40,000 molecules per haploid yeast cell, it has been proposed that Mcm10 not only has roles in DNA replication and heterochromatin formation, but also serves as a facilitator of other protein interactions. Using this system, it was shown that Mcm10 may participate in several ternary protein complexes.
Apger, Jennifer. (January 2010). Interaction and Functional Analysis of Drosophila Mcm10 (Master's Thesis, East Carolina University). Retrieved from the Scholarship. (http://hdl.handle.net/10342/2873.)
Apger, Jennifer. Interaction and Functional Analysis of Drosophila Mcm10. Master's Thesis. East Carolina University, January 2010. The Scholarship. http://hdl.handle.net/10342/2873. October 27, 2020.
Apger, Jennifer, “Interaction and Functional Analysis of Drosophila Mcm10” (Master's Thesis., East Carolina University, January 2010).
Apger, Jennifer. Interaction and Functional Analysis of Drosophila Mcm10 [Master's Thesis]. Greenville, NC: East Carolina University; January 2010.
East Carolina University
Showing items related by title, author, creator and subject.
Phylogenetics and Systematics of the Millipede genus Brachycybe Wood, 1864 (Platydesmida: Andrognathidae) Spruill, Chad Lee (East Carolina University, 2010)The genus Brachycybe Wood, 1864 (Platydesmida: Andrognathidae) belongs to an ancient millipede lineage that has persisted since the Miocene. The group displays a Holarctic distribution with species in the eastern Nearctic, ...
The bhuTUV and bhuO genes play vital roles in the ability of Brucella abortus to use heme as an iron source and are regulated in an iron-responsive manner by RirA and Irr Ojeda, Jenifer F. (East Carolina University, 2012)Brucella abortus is a Gram negative intracellular pathogen that causes the zoonotic disease brucellosis. Antibiotic treatment for brucellosis in humans is prolonged and sometimes followed by relapses. Currently, the United ...
The Role of eIF4G-1 Isoforms and Live Cell Imaging of IRES-mediated Translation Initiation in Breast Cancer Cells Crosson, Whitney K. (East Carolina University, 2012)The development of cancer is a consequence of mutations that lead to dysfunctional cell processes such as unrestrained cell proliferation, resistance to apoptosis, and improper regulation of cell processes such as translation. ...