Identification and Characterization of Proteins for the Initiation of DNA Replication in Saccharomyces Cerevisiae PDF Download
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Author: Tsz Choi Chan
Publisher:
ISBN:
Category : DNA replication
Languages : en
Pages : 244
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Book Description
Author: Tsz Choi Chan
Publisher:
ISBN:
Category : DNA replication
Languages : en
Pages : 244
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Book Description
Author: Yuanliang Zhai
Publisher:
ISBN:
Category : DNA replication
Languages : en
Pages : 189
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Book Description
Author: Lin Huo
Publisher:
ISBN:
Category : DNA replication
Languages : en
Pages : 334
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Book Description
Author: John A. Bryant
Publisher: CRC Press
ISBN: 1351088416
Category : Science
Languages : en
Pages : 477
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Book Description
This texts discusses DNA replication in plants including chapters on; functional chromosomal structure, the biochemistry of DNA replication, Control of DNA replication, Replication of plant organelle DNA, replication of DNA viruses in plants, and DNA damage, repair, and mutagenesis.
Author: Rohan Dhiraj Gidvani
Publisher:
ISBN:
Category :
Languages : en
Pages : 244
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Book Description
A crucial step in eukaryotic cell proliferation is the initiation of DNA replication, a tightly regulated process mediated by a multitude of protein factors. In Saccharomyces cerevisiae, this occurs as a result of the concerted action of an assembly of proteins acting at origins of replication, known as the pre-replicative complex (pre-RC). While many of the mechanisms pertaining to the functions of these proteins and the associations amongst them have been explored experimentally, mathematical models are needed to effectively explore the network's dynamic behaviour. An ordinary differential equation (ODE)-based model of the protein-protein interaction network describing DNA replication initiation was constructed. The model was validated against quantified levels of protein factors determined in vivo and from the literature over a range of cell cycle timepoints. The model behaviour conforms to perturbation trials previously reported in the literature and accurately predicts the results of knockdown experiments performed herein. Furthermore, the DNA replication model was successfully incorporated into an established model of the entire yeast cell cycle, thus providing a comprehensive description of these processes. A screen for novel DNA damage response proteins was investigated using a unique proteomics approach that uses chromatin fractionation samples to enrich for factors bound to the DNA. This form of sub-cellular fractionation was combined with differential-in-gel-electrophoresis (DIGE) to detect and quantify low abundance chromatin proteins in the budding yeast proteome. The method was applied to analyze the effect of the DNA damaging agent methyl methanesulfonate (MMS) on levels of chromatin-associated proteins. Up-regulation of several previously characterized DNA damage checkpoint-regulated proteins, such as Rnr4, Rpa1 and Rpa2, was observed. In addition, several novel DNA damage responsive proteins were identified and assessed for genotoxic sensitivity. A strain in which the expression of the Ran-GTPase binding protein Yrb1 was reduced was found to be hypersensitive to genotoxic stress, pointing to a role for this nuclear import-associated protein in DNA damage response. The model presented in this thesis provides a tool for exploring the biochemical network of DNA replication. This is germane to the exploration of new cancer therapeutics considering the link between this disease (and others) and errors in proper cell cycle regulation. The high functional conservation between cell cycle mechanisms in humans and yeast allows predictive analyses of the model to be extrapolated towards understanding aberrant human cell proliferation. Importantly, the model is useful in identifying potential targets for cancer treatment and provides insights into developing highly specific anti-cancer drugs. Finally, the characterization of factors in the proteomic screen opens the door to further investigation of the roles of potential DNA damage response proteins.
Author: Yun Jiang
Publisher:
ISBN:
Category :
Languages : en
Pages : 336
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Author: Chia-Lin Wei (Ph.D. in microbiology)
Publisher:
ISBN:
Category :
Languages : en
Pages : 422
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Author: Mami Kainuma
Publisher:
ISBN:
Category :
Languages : en
Pages : 326
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Author: Su-Wen Ho
Publisher:
ISBN:
Category : Electronic dissertations
Languages : en
Pages : 157
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Book Description
Gene expression is an elaborate and finely tuned process involving the regulated interactions of multiple proteins with promoter and enhancer elements. A variety of approaches are currently used to study these interactions in vivo, in vitro as well as in silico. With the genome sequences of many organisms now readily available, a plethora of DNA functional elements have been predicted, but the process of identifying the proteins that bind to them in vivo remains a bottleneck. I developed two high-throughput assays to address this issue. The first is a modification of the yeast "one-hybrid" assay. The second is probing protein microarrays with DNA sequence elements. Using these methods, I identified two proteins, Sef1 and Yjl103c, that bind to the same DNA sequence element. Sef1 and Yjl103c are little-characterized members of the zinc cluster family of transcription factors of S. cerevisiae. Characterization of their mechanism of action as well as identification of some of their target genes leads to the conclusion that they play a pivotal role in the transcriptional regulation of utilization of nonfermentable carbon sources by budding yeast.
Author: S. Sweet
Publisher:
ISBN:
Category :
Languages : en
Pages :
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Book Description
The initiation of DNA replication at the onset of S phase in eukaryotic cells is a critically important and tightly regulated process. Multiple origins of replication in the genome must be co-ordinately regulated such that duplication of the chromosomes is complete before cell division, whilst also ensuring that no sections of the DNA are over-replicated. In G1 phase of the cell cycle, a large 'pre-replicative complex' (pre-RC) forms at origins consisting of a hexameric Origin Recognition Complex (ORC) as well as Cdc6, Cdt1 and another hexameric complex known as the Minichromosome Maintenance (MCM) complex. At the onset of S phase, two cell cycle regulated protein kinases, the Cyclin Dependent Kinase (CDK) and Cdc7, are activated. Phosphorylation of various proteins by these two enzymes triggers formation of large 'replisome' complexes, initiation of DNA replication from each origin, and disassembly of the pre-RCs. Pre-RC re-assembly is subsequently inhibited until kinase activity falls again after cell division. In this study, we have set about identifying substrates of both CDK and Cdc7 involved in DNA replication in the budding yeast Saccharomyces cerevisiae. Two techniques are employed, the in vitro phosphorylation of arrays of peptides and phosphorylation of pre-RCs assembled in cell-free yeast extracts. Peptide arrays provide a high throughput technique for screening large numbers of potential substrates in a single experiment, whilst pre-RC phosphorylation allows consideration of both tertiary and quaternary structures of the in vivo kinase substrate. Several potential novel substrates of both CDK and Cdc7 are revealed. Pre-RC phosphorylation also reveals a previously unreported phosphorylation of Orc1 by a third kinase which has been identified as Casein Kinase II (CKII).
