Author: Parie Garg
Publisher:
ISBN:
Category :
Languages : en
Pages : 242

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Author: Giovanni Maga
Publisher: World Scientific
ISBN: 9813226420
Category : Science
Languages : en
Pages : 398

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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.

Author: Joseph L. Stodola
Publisher:
ISBN:
Category : Electronic dissertations
Languages : en
Pages : 178

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The work described in this dissertation focuses on several aspects of DNA replication in the model organism Saccharomyces cerevisiae, with particular attention paid to the function of the replicative DNA polymerase [delta] (Pol [delta]), and its functions in Okazaki fragment synthesis and maturation. The first major theme of this dissertation is investigating the role that metal binding motifs play in the structure and function of Pol [delta] and other budding yeast polymerases. First, I discuss the role that two metal binding motifs within the catalytic subunit of Pol [delta] play in creating the multi-subunit polymerase complex and in promoting crucial interactions with the replication sliding clamp, proliferating cell nuclear antigen (PCNA). Next, I describe work defining the importance of similar metal binding motifs in the translesion DNA polymerase [zeta] (Pol [zeta]). This yielded the observation that the two accessory subunits of Pol [delta], Pol31 and Pol32, are also constitutive members of a four-subunit Pol [zeta] complex. Finally, I describe the creation of a chimeric DNA polymerase comprising the bacteriophage RB69 DNA polymerase fused to the metal binding domain of the Pol [delta] catalytic subunit. We show that this chimeric polymerase can form a multimeric complex containing the Pol [delta] accessory subunits, interact with PCNA, and support DNA replication in vivo. This data provided insight into the structural requirements of the lagging strand replication machinery. The second major theme is Pol [delta]'s crucial role in synthesizing Okazaki fragments and participating in the removal of initiator RNA, called Okazaki fragment maturation. I first describe my work developing a system to study the activity of Pol [delta] in higher kinetic detail than previous studies, using rapid-quench techniques. This work yielded insights into how Pol [delta] performs DNA synthesis and strand displacement synthesis, as well as accomplishes nick translation, requiring collaboration between Pol [delta] and the flap-endonuclease FEN1. The next chapter describes the production and characterization of engineered PCNA heterotrimers. These proteins were produced to test the 'toolbelt' model, which is the hypothesis that PCNA binds multiple enzymes simultaneously to increase the efficiency of DNA metabolism processes involving multiple enzymes. Finally, there has been a growing interest among those studying lagging strand synthesis into how potential impediments to the Okazaki fragment maturation machinery are resolved; we show that although the transcription factor Rap1 can block strand displacement synthesis by Pol [delta] when it is bound to DNA, this block can be resolved through the action of the helicase Pif1. In sum, these studies provide insight into how Pol [delta]'s structure dictates its function, as well as addresses larger mechanistic questions concerning how lagging strand DNA replication is accomplished.

Author: Min You
Publisher:
ISBN:
Category : DNA polymerases
Languages : en
Pages : 186

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Author: Katsuhiko S. Murakami
Publisher: Springer Science & Business Media
ISBN: 3642397964
Category : Science
Languages : en
Pages : 342

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This book provides a review of the multitude of nucleic acid polymerases, including DNA and RNA polymerases from Archea, Bacteria and Eukaryota, mitochondrial and viral polymerases, and other specialized polymerases such as telomerase, template-independent terminal nucleotidyl transferase and RNA self-replication ribozyme. Although many books cover several different types of polymerases, no book so far has attempted to catalog all nucleic acid polymerases. The goal of this book is to be the top reference work for postgraduate students, postdocs, and principle investigators who study polymerases of all varieties. In other words, this book is for polymerase fans by polymerase fans. Nucleic acid polymerases play a fundamental role in genome replication, maintenance, gene expression and regulation. Throughout evolution these enzymes have been pivotal in transforming life towards RNA self-replicating systems as well as into more stable DNA genomes. These enzymes are generally extremely efficient and accurate in RNA transcription and DNA replication and share common kinetic and structural features. How catalysis can be so amazingly fast without loss of specificity is a question that has intrigued researchers for over 60 years. Certain specialized polymerases that play a critical role in cellular metabolism are used for diverse biotechnological applications and are therefore an essential tool for research.

Author: Kathleen M. Downey
Publisher:
ISBN:
Category :
Languages : en
Pages : 6

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Author: Tianli Xia
Publisher:
ISBN:
Category : DNA polymerases
Languages : en
Pages : 202

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Author: Franklin Bueno
Publisher:
ISBN:
Category :
Languages : en
Pages : 93

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Author: Robert Vincent Talanian
Publisher:
ISBN:
Category :
Languages : en
Pages : 328

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Author: Xiangyang Li
Publisher:
ISBN:
Category :
Languages : en
Pages : 306

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