Advanced search options

Advanced Search Options 🞨

Browse by author name (“Author name starts with…”).

Find ETDs with:

in
/  
in
/  
in
/  
in

Written in Published in Earliest date Latest date

Sorted by

Results per page:

You searched for id:"oai:etd.ohiolink.edu:csu1504869235809659". One record found.

Search Limiters

Last 2 Years | English Only

No search limiters apply to these results.

▼ Search Limiters

1. Dasari, Anvesh. Using Nucleotide Analogs as Biochemical Probes to Evaluate the Mechanisms Involved in Translesion Replication by a High Fidelity DNA Polymerase.

Degree: PhD, College of Sciences and Health Professions, 2017, Cleveland State University

Translesion DNA synthesis (TLS) allows DNA polymerases to incorporate nucleotides opposite and beyond damaged DNA. This activity is an important risk factor for the initiation and progression of genetic diseases including cancer. My study evaluates the ability of a high-fidelity DNA polymerase to perform TLS with 8-oxo-guanine, a pro-mutagenic DNA lesion formed by reactive oxygen species. Using modified purine and non-natural indole analogs as biochemical probes, I have evaluated the influence of desolvation, hydrogen bonding interactions, and shape complementarity towards nucleotide binding and incorporation opposite the miscoding lesion 8-oxo-guanine by the high fidelity gp43exo- DNA polymerase. In Chapter II, I used modified purine nucleotide analogs to provide evidence that nucleobase desolvation and hydrogen bonding interactions play a crucial role towards binding and incorporation opposite 8-oxo-guanine. This was further confirmed by studies in Chapter III through kinetic characterization using non-natural indole nucleotide analogs. Overall, I have demonstrated that the binding affinity of the incoming dNTP is controlled by the overall hydrophobicity of the nucleobase. However, the rate constant for the conformational change preceding chemistry is regulated by hydrogen-bonding interactions and play a much larger role during the replication of miscoding lesions such as 8-oxo-G. Results generated here for the replication of 8-oxo-guanine were compared to those published for the replication of an abasic site, a non-instructional DNA lesion. With both lesions, nucleobase hydrophobicity is a common feature that controls nucleotide binding whereas the physical nature of the lesion, i.e., miscoding versus non-instructional, influences the rate constant of the conformational change step that precedes phosphoryl transfer. Collectively, these studies highlight the importance of nucleobase desolvation as a key physical feature that can hinder or facilitate the misreplication of structurally diverse DNA lesions. In Chapter IV, I have investigated the unique selectivity of a modified purine analog to DNA polymerases with varied biological function. Active analysis demonstrated that differential architecture of high and low fidelity DNA polymerases plays a predominant role in the replication of damaged DNA. Therefore, this differential utilization provides a unique opportunity to develop a chemical probe to monitor translesion DNA synthesis in-vivo. Advisors/Committee Members: Berdis, Anthony (Committee Chair).

Subjects/Keywords: Analytical Chemistry; Biochemistry; Chemistry

Record DetailsSimilar RecordsGoogle PlusoneFacebookTwitterCiteULikeMendeleyreddit

APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Dasari, A. (2017). Using Nucleotide Analogs as Biochemical Probes to Evaluate the Mechanisms Involved in Translesion Replication by a High Fidelity DNA Polymerase. (Doctoral Dissertation). Cleveland State University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=csu1504869235809659

Chicago Manual of Style (16th Edition):

Dasari, Anvesh. “Using Nucleotide Analogs as Biochemical Probes to Evaluate the Mechanisms Involved in Translesion Replication by a High Fidelity DNA Polymerase.” 2017. Doctoral Dissertation, Cleveland State University. Accessed November 19, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=csu1504869235809659.

MLA Handbook (7th Edition):

Dasari, Anvesh. “Using Nucleotide Analogs as Biochemical Probes to Evaluate the Mechanisms Involved in Translesion Replication by a High Fidelity DNA Polymerase.” 2017. Web. 19 Nov 2017.

Vancouver:

Dasari A. Using Nucleotide Analogs as Biochemical Probes to Evaluate the Mechanisms Involved in Translesion Replication by a High Fidelity DNA Polymerase. [Internet] [Doctoral dissertation]. Cleveland State University; 2017. [cited 2017 Nov 19]. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=csu1504869235809659.

Council of Science Editors:

Dasari A. Using Nucleotide Analogs as Biochemical Probes to Evaluate the Mechanisms Involved in Translesion Replication by a High Fidelity DNA Polymerase. [Doctoral Dissertation]. Cleveland State University; 2017. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=csu1504869235809659

.