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Probing the Chemistry and Enzymology of Translesion DNA Synthesis: Applications in Developing a Novel “Theranostic” Agent against Leukemia

Motea, Edward A.

Abstract Details

2012, Doctor of Philosophy, Case Western Reserve University, Chemistry.
The replication and maintenance of the genetic information stored in DNA are required for all forms of life. As such, any inappropriate alteration to the information programmed into the nucleotide sequence of cellular DNA could interfere with normal DNA replication, and thus lead to cell death. Indeed, high doses of chemotherapeutic agents and/or ionizing radiation are commonly used in advanced cancer patients to create toxic DNA lesions in malignant cells. However, these conventional DNA-damaging agents rarely eradicate cancer cells completely due to their promutagenic nature. Translesion synthesis (TLS), the replication of damaged DNA, provides the molecular mechanism of DNA synthesis in cancer cells following chemotherapy that ultimately leads to drug resistance and/or the initiation of secondary malignancies. Thus, inhibition of TLS could sensitize cancer cells to the effects of chemotherapy, and prevent the emergence of cancer drug resistance and carcinogenesis. Towards this goal, I have investigated the enzymology and chemistry of a biological phenomenon known as the “A-Rule” of TLS which refers to the ability of various replicative DNA polymerases to preferentially incorporate 2’-deoxyadenosine nucleotide opposite non-instructional DNA lesions such as the abasic site, blunt-ended DNA, and 5’ to 3’ single-stranded DNA overhang. Using a series of non-natural nucleotides that I have designed and synthesized as probes, I was able to determine crucial biophysical features of the incoming nucleotides that facilitate various DNA polymerases to preferentially and efficiently catalyze its incorporation opposite these lesions. The wealth of information from these studies has led to the development of a “clickable” non-natural nucleotide that is selectively incorporated opposite non-instructional lesions, and then terminates DNA replication, repair and recombination. I have also demonstrated that the corresponding nucleoside has combined therapeutic and diagnostic activities against leukemia. This occurs as a result of its metabolism and incorporation in the DNA that could be detected via “click” chemistry with a fluorogenic reporter. Collectively, the mechanistic insights gained from this study has important implications beneficial to the rational design of novel molecules as probes for biochemical study, for the expansion of the genetic alphabet or as therapeutic agent.
Irene Lee (Advisor)
Thomas Gray (Committee Chair)
Thomas Gerken (Committee Member)
Yanming Wang (Committee Member)
Anthony Berdis (Committee Member)
333 p.

Recommended Citations

Citations

  • Motea, E. A. (2012). Probing the Chemistry and Enzymology of Translesion DNA Synthesis: Applications in Developing a Novel “Theranostic” Agent against Leukemia [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1323382718

    APA Style (7th edition)

  • Motea, Edward. Probing the Chemistry and Enzymology of Translesion DNA Synthesis: Applications in Developing a Novel “Theranostic” Agent against Leukemia. 2012. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1323382718.

    MLA Style (8th edition)

  • Motea, Edward. "Probing the Chemistry and Enzymology of Translesion DNA Synthesis: Applications in Developing a Novel “Theranostic” Agent against Leukemia." Doctoral dissertation, Case Western Reserve University, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=case1323382718

    Chicago Manual of Style (17th edition)