Reconstitution and Characterization of Human DNA Polymerase Delta Four Subunit Holoenzyme and Proteomic Analysis of DNA Polymerase Delta Complex PDF Download
Are you looking for read ebook online? Search for your book and save it on your Kindle device, PC, phones or tablets. Download Reconstitution and Characterization of Human DNA Polymerase Delta Four Subunit Holoenzyme and Proteomic Analysis of DNA Polymerase Delta Complex PDF full book. Access full book title Reconstitution and Characterization of Human DNA Polymerase Delta Four Subunit Holoenzyme and Proteomic Analysis of DNA Polymerase Delta Complex by Nayef Ali Mazloum. Download full books in PDF and EPUB format.
Author: Nayef Ali Mazloum
Publisher:
ISBN:
Category :
Languages : en
Pages : 162
Get Book Here
Book Description
Author: Nayef Ali Mazloum
Publisher:
ISBN:
Category :
Languages : en
Pages : 162
Get Book Here
Book Description
Author: Giovanni Maga
Publisher: World Scientific
ISBN: 9813226420
Category : Science
Languages : en
Pages : 398
Get Book Here
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.
Author: Zachary Forest Pursell
Publisher:
ISBN:
Category :
Languages : en
Pages : 294
Get Book Here
Book Description
Author: Ulrich Hbscher
Publisher: World Scientific
ISBN: 9814299162
Category : Science
Languages : en
Pages : 338
Get Book Here
Book Description
Maintenance of the information embedded in the genomic DNA sequence is essential for life. DNA polymerases play pivotal roles in the complex physiological processes of DNA replication and repair. Besides the tasks in vivo, DNA polymerases are the workhorses in numerous biotechniques such as polymerase chain reaction (PCR), cDNA cloning, genome sequencing, nucleic acids–based diagnostics, as well as techniques to analyze ancient and otherwise damaged DNA. The authors have recently witnessed the discovery of a plethora of novel DNA polymerases with specialized properties whose physiological functions are only just beginning to be understood. This book summarizes the current knowledge of these fascinating enzymes in viruses, bacteria, archaea and eukaryotes. Moreover, some diseases are related to DNA polymerase defects, and chemotherapy through inhibition of DNA polymerases is used to fight HIV, Herpes, as well as Hepatitis B and C infections. This book will appeal to a broad audience including basic scientists, diagnostic laboratories, and clinicians who will gain an invaluable understanding of these fascinating enzymes.
![Identification and Characterization of a Novel Protein that Interacts with the 50-kDa Subunit of Human DNA Polymerase [delta] and PCNA PDF](https://books.google.com/books/content/images/frontcover/lkr1tgAACAAJ?fife=w80-h100)
Author: Hua He
Publisher:
ISBN:
Category : DNA polymerases
Languages : en
Pages : 214
Get Book Here
Book Description
Author: Alexander Rodney Abbas
Publisher:
ISBN:
Category :
Languages : en
Pages : 316
Get Book Here
Book Description
Author: Cindy M. Jaime
Publisher:
ISBN:
Category :
Languages : en
Pages : 69
Get Book Here
Book Description
![Purification and Characterization of Recombinant Human DNA Polymerase [delta] Expressed in Baculovirus-infected Insect Cells PDF](https://books.google.com/books/content/images/frontcover/j3rGNwAACAAJ?fife=w80-h100)
Author: Jin-Qiu Zhou
Publisher:
ISBN:
Category : DNA polymerases
Languages : en
Pages : 180
Get Book Here
Book Description
Author: Joseph Cardina
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
Get Book Here
Book Description
In lagging strand DNA synthesis, DNA polymerase [delta] (pol [delta]) holoenzymes composed of pol [delta] and the proliferating cell nuclear antigen (PCNA) sliding clamp perform high fidelity DNA synthesis with little error. The pol [delta] holoenzyme occasionally encounters structural deformities in the native DNA sequence called DNA lesions. Upon encountering these lesions, it was previously believed that pol [delta] would stall on the DNA template at some point before the lesion launching a DNA damage tolerance (DDT) pathway to replicate the lesion and remaining DNA before allowing pol [delta] to resume its typical high fidelity DNA synthesis. Recent literature within the past 20 years or so has challenged this perspective by showing human pol [delta] can replicate various DNA lesions. These studies have questioned the role that DDT plays in lagging strand DNA synthesis upon encounter of a DNA lesion, thus providing a need for further analysis. As a result, we aimed to quantitatively characterize the encounters of pol [delta] with several biologically relevant DNA lesions at physiological conditions. Our results show that pol [delta] supports dNTP incorporation while inserting across from, extending from, and elongating past several different DNA lesions, and the exact extent of these values is dependent on the exonuclease activity of pol [delta] and the identity of the lesion itself. We also found that of the pol [delta] that dissociates upon encountering the lesion, the dissociation events are unevenly distributed around the lesion with different distributions depending on the lesion. Our findings show that lagging strand DNA synthesis is more complex than previously thought while advancing our understanding of the role of DDT in lagging strand DNA synthesis.
Author: Nikunj Bhatt
Publisher:
ISBN:
Category : DNA polymerases
Languages : en
Pages : 96
Get Book Here
Book Description
Abstract: Since 1999, the human genome project has led to the discovery of several novel DNA polymerases. Among these new polymerases were human DNA polymerase ?. (Pol?) and Sulfolobus solfataricus DNA polymerase IV (Dpo4). PoP, an X family polymerase, displays both 5'-2-deoxyribose-5-phosphate lyase (dRPase) activity and polymerase activity and efficiently incorporates nucleotides into short-gapped primer- primer/templates, the natural substrate for short-patch base excision repair (BER). Crystal studies of the ternary structure of truncated PoIA (tPolX) reveal that a 5'- phosphate of a downstream primer interacts with a positively-charged pocket in the dRPase domain. In this study, we constructed three substrates: (i) a 21-19/41-mer single nucleotide gapped DNA with a 5'-phosphate on the terminal end of the downstream 19- mer; (ii) a 21-19/41-mer single nucleotide gapped DNA with a 5'-OH on the downstream 19-mer; and (iii) a 21/41-mer with no downstream primer. Pre-steady state kinetics revealed that Pol[lamda]'s incorporation efficiency for the DNA substrate with the 5'-phosphate moiety on the downstream 1 9-mer primer is 11-fold more efficient at incorporating a correct nucleotide compared to the DNA substrate with the 5'-OH on the terminal end of the downstream 19-mer, and 160-fold more efficient than the DNA substrate with no downstream primer. Another aim of this thesis was to characterize Po1[lamda]' s preference for deoxynucleotides over ribonucleotides via pre-steady state kinetics. A previous study with polymerase u (Polu) revealed that the glycine433 residue mutated to tyrosine dramatically increased sugar selectivity for that enzyme. Sequence alignment of the M a-helix of Po1[lamda] and three other X family polymerases, Polu, terminal deoxynucleotidyl transferase (TDT), and polymerase [beta] (Polf[beta]), suggested that its tyrosine505 may be involved in determining sugar selectivity, while the crystal structure of the tPo[lamda]. ternary complex suggested that the backbone of the M a-helix blocks the 2'-OH of ribonucleotides forming a "stearic gate". In this thesis, the tyrosine505 mutated to glycine actually increased sugar selectivity indirectly supporting the stearic gate hypothesis. The second enzyme studied, Dpo4, a Y family polymerase that bypasses lesions and exhibits low fidelity, consists of an extra little finger domain in addition to the standard catalytic core, consisting of the finger, thumb, and palm domains. The little finger, which has been shown to be important to polymerase activity, is tethered to the thumb domain via 14 amino acid linker (P1 = 10), known as the little finger linker. Using 1, 4, and 6 glycine additions and 1, 4, and 6 deletions in the center of the little finger linker, a fluorescent titration assay revealed a significant decrease in binding affinity as the size of the little finger linker was increased and decreased. This suggests nature has optimized the length of the little finger linker. In addition, a kink in the little finger linker was removed by a proline236 substitution to alanine which did not significantly effect DNA binding to Dpo4. Finally, an optimized procedure for the cisplatination of an 1 8-mer DNA substrate was prepared and described in this thesis.
