Kinetic Characterizations of DNA Replication Fidelity of E. Coli DNA Polymerase II PDF Download
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Author: Zhijie Wang
Publisher:
ISBN:
Category :
Languages : en
Pages : 212
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Book Description
Author: Zhijie Wang
Publisher:
ISBN:
Category :
Languages : en
Pages : 212
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Book Description
Author: Walter J. Zahurancik
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages :
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Book Description
The results of my work lay the foundation for future studies exploring the effects of other DNA replication enzymes on the polymerization and fidelity of hPole. Furthermore, my work provides the basis for investigating the mechanism of mutagenesis in tumors containing hPole exonuclease domain mutants. Overall, this study represents an important advance toward our comprehensive understanding of human DNA replication.
Author: Errol C. Friedberg
Publisher: American Society for Microbiology Press
ISBN: 1555813194
Category : Science
Languages : en
Pages : 2587
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Book Description
An essential resource for all scientists researching cellular responses to DNA damage. • Introduces important new material reflective of the major changes and developments that have occurred in the field over the last decade. • Discussed the field within a strong historical framework, and all aspects of biological responses to DNA damage are detailed. • Provides information on covering sources and consequences of DNA damage; correcting altered bases in DNA: DNA repair; DNA damage tolerance and mutagenesis; regulatory responses to DNA damage in eukaryotes; and disease states associated with defective biological responses to DNA damage.
Author: Merilyn Blair-Johnson
Publisher:
ISBN:
Category : DNA polymerases
Languages : en
Pages : 338
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Book Description
Author: David Lewis Sloane
Publisher:
ISBN:
Category :
Languages : en
Pages : 294
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Book Description
Author: Judith L. Campbell
Publisher: Academic Press
ISBN:
Category : Nature
Languages : en
Pages : 770
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Book Description
The critically acclaimed laboratory standard for forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerlyawaited, frequently consulted, and praised by researchers and reviewers alike. More than 250 volumes have been published (all of them still in print) and much of the material is relevant even today--truly an essential publication for researchers in all fields of life sciences. Key Features * Includes descriptions of functional, structural, kinetic, and genetic methods for analyzing major enzymes of DNA replication * Describes strategies for studying interactions of these proteins during replication * Provides comprehensive descriptions of uses of prokaryotic and eukaryotic crude in vitro replication systems and reconstitution of such systems from purified proteins * Includes methods for analyzing DNA replication in vivo
Author: Christopher K. Mathews
Publisher:
ISBN:
Category : Science
Languages : en
Pages : 432
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Book Description
Author: David Stuart
Publisher: BoD – Books on Demand
ISBN: 953510991X
Category : Medical
Languages : en
Pages : 502
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Book Description
DNA replication is a fundamental part of the life cycle of all organisms. Not surprisingly many aspects of this process display profound conservation across organisms in all domains of life. The chapters in this volume outline and review the current state of knowledge on several key aspects of the DNA replication process. This is a critical process in both normal growth and development and in relation to a broad variety of pathological conditions including cancer. The reader will be provided with new insights into the initiation, regulation, and progression of DNA replication as well as a collection of thought provoking questions and summaries to direct future investigations.
Author: Tyler Lane Dangerfield
Publisher:
ISBN:
Category :
Languages : en
Pages : 532
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Book Description
Enzymes that catalyze DNA and RNA replication are biologically important and provide examples to understand enzyme specificity as the correct substrate and alternate substrates are well known. The DNA polymerase from bacteriophage T7 has been a model system for understanding high-fidelity DNA replication, yet outstanding questions remain about the controversial nucleotide induced conformational change. I developed methods to directly measure the conformational change by site specifically incorporating a fluorescent unnatural amino acid into the enzyme at a position that gives a fluorescence signal upon nucleotide binding. In addition, I developed high throughput methods to analyze kinetic samples by capillary electrophoresis to assist in the kinetic analysis of DNA replication. Kinetic studies show that the conformational change is the primary determinant of specificity at the polymerase active site. I also characterized the enzyme’s 3’--5’ proofreading exonuclease activity and showed that the enzyme has multiple opportunities to correct mistakes during replication. I also used molecular dynamics simulation methods to propose a structure of the proofreading exonuclease editing complex. While finishing studies on the T7 DNA polymerase, the SARS-CoV-2 pandemic devastated the world. I used techniques developed in my previous studies to characterize the RNA dependent RNA polymerase (RdRp) from SARS-CoV-2, a prime target for antiviral drugs to stop replication of the virus. I developed methods to express the tag free complex in E. coli and subsequent purification to give highly active enzyme, which for the first time met standards for physiologically relevant activity. I characterized the pre-steady state kinetics of UTP and ATP incorporation and found that the enzyme catalyzes nucleotide incorporation at a fast rate, sufficient to replicate the 30 kb viral genome in less than 2 minutes. The ATP analog Remdesivir triphosphate, the only FDA approved treatment for COVID-19, was then studied to calculate the discrimination relative to ATP. We found that Remdesivir is incorporated more efficiently than ATP and acts as a delayed chain terminator, although this termination can be partially overcome by incubation with high nucleotide concentrations. Cryo-EM was used to determine the structure of the Remdesivir stalled RdRp complex and we found that the translocation step was inhibited after incorporation of 3 Remdesivirs due to a steric block between Remdesivir and the enzyme
Author: Giovanni Maga
Publisher: World Scientific
ISBN: 9813226420
Category : Science
Languages : en
Pages : 398
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Book Description
Maintenance of the information embedded in the genomic DNA sequence is essential for life. DNA polymerases play pivotal roles in the complex processes that maintain genetic integrity. Besides their tasks in vivo, DNA polymerases are the workhorses in numerous biotechnology applications such as the polymerase chain reaction (PCR), cDNA cloning, next generation sequencing, nucleic acids based diagnostics and in techniques to analyze ancient and otherwise damaged DNA (e.g. for forensic applications). Moreover, some diseases are related to DNA polymerase defects and chemotherapy through inhibition of DNA polymerases is used to fight HIV, Herpes and Hepatitis B and C infections. This book focuses on (i) biology of DNA polymerases, (ii) medical aspects of DNA polymerases and (iii) biotechnological applications of DNA polymerases. It is intended for a wide audience from basic scientists, to diagnostic laboratories, to companies and to clinicians, who seek a better understanding and the practical use of these fascinating enzymes.
