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Expression, purification and characterisation of the human papillomavirus type 2aE1 proteinLloansiÌ Vila, Ariadna January 2002 (has links)
No description available.
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Investigations of the origins of replication of herpesvirus saimiriSchofield, Andrea January 1994 (has links)
No description available.
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Characterisation of Staphylococcus aureus GyrB and ParE proteins and their interactions with antibacterial agentsSayer, Penelope Jane January 1998 (has links)
No description available.
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Structural investigations of the Bacillus subtilis SPP1 phage G39P helicase inhibitor loading proteinBailey, Scott January 2002 (has links)
No description available.
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Properties and applications of a mutagenic nucleoside analogue dPHarris, Victoria Helen January 2001 (has links)
No description available.
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Studies on DNA mismatch repair in Pseudomonas aeruginosaGibson, Roderick J. January 1992 (has links)
No description available.
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Analysis of the cdc21'+ gene of Schizosaccharomyces pombeCoxon, Angela January 1992 (has links)
No description available.
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Mathematical models for DNA replication machineryHameister, Heike January 2012 (has links)
DNA replication and associated processes take place in all living organisms with the same constitutions. The knowledge of the duplication process, chromatin building and repair mechanisms has increased explosively over the last years, but the complex interplay of different proteins and their mechanisms are not conceived properly. During DNA replication, the DNA has to be unpacked, duplicated and finally repacked into chromatin. These steps require different proteins, e.g. new histone proteins on demand to secure an error-free and undelayed DNA replication. This thesis includes different mathematical models for DNA replication, repair and chromatin formation, which are based on experimental results. Three models of chromatin formation provide a simplified description of histone gene expression and protein synthesis during G1/S/G2 phase and include the contribution of different regulatory elements. Furthermore, all models present two different mechanisms of regulation to test possible scenarios of newly synthesised histones and free DNA binding sites. The basic model presents a single histone gene, which codes for a single histone protein. The stem-loop binding protein (SLBP) acts as a master regulator, which is only present during S phase. Different analyses of early S-phase, over- and underexpressed replication and the down-regulation of SLBP proof the model under extreme conditions. This basic model serves as a template for further scenarios with several genes and different histone families. For this, a second model is realised to simulate imbalances in the histone mRNA synthesis and translation. Additionally, a third model tests a gene knock-out and mRNA silencing. The initial histone model is able to qualitatively reproduce experimental observations and shows basic regulatory principles. The adaptation with several genes and different histone families presents qualitatively different system responses for the discussed regulatory mechanisms and illustrates the ability to compensate the effect of mRNA silencing.
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Protein-protein interactions involved in baculovirus DNA replicationEvans, Jay T. 17 February 1998 (has links)
The yeast two-hybrid system was used to examine
interactions between the nine proteins involved in baculovirus DNA
replication. From the six proteins required for DNA replication,
four protein-protein interactions were identified, including an
interaction between LEF-1 and LEF-2, LEF-3 and itself, LEF-3 and
P143 (Helicase), and an interaction between IE-1 and itself.
The replication factors LEF-1 and LEF-2 interacted in both
yeast two-hybrid assays and glutathione S-transferase fusion
affinity assays. Using the yeast two-hybrid system, we mapped
the interaction domain of LEF-2 to amino acids between positions
20 and 60. Deletion analysis of LEF-1 failed to reveal an
interaction domain, suggesting that either multiple interaction
domains exists or the deletions disrupted secondary structures
required for the interaction. All of the deletions which were
unable to interact also failed to support significant levels of
transient DNA replication, suggesting that this interaction plays a
significant role in DNA replication.
The baculovirus single-stranded DNA binding protein, LEF-3, interacts with itself in yeast two-hybrid assays and glutathione S-transferase fusion affinity assays. Deletions of LEF-3, which were unable to interact with full length LEF-3, also failed to support transient DNA replication, suggesting that this interaction is required for the proper function of LEF-3. LEF-3 was purified to apparent homogeneity and analyzed by analytical ultracentrifugation, native PAGE and MALDI mass spectrometry, identifying the oligomeric structure of LEF-3 as a homotrimer.
In addition to interacting with itself, LEF-3 also interacts with P143 in yeast two-hybrid assays, immunoprecipitation experiments, and co-purification from a single-stranded DNA agarose column. The yeast two-hybrid system was used to map the LEF-3 interaction domain to the N-terminal 165 amino acids of LEF-3. Deletion analysis of P143 failed to reveal a delimited interaction domain. C-terminal deletions of LEF-3 containing amino acids 1 to 165 were unable to interact with full length LEF-3, indicating that the interaction of LEF-3 with itself (trimerization) is not required for the interaction between LEF-3 with P143. / Graduation date: 1998
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The Roles of Conserved Dbf4 Motifs in DNA Replication and Checkpoint Responses in Saccharomyces cerevisiaePrasad, Ajai Anand 23 December 2009 (has links)
The Dbf4 protein is involved in the initiation of DNA replication, in complex with Cdc7 kinase, and also plays a role in the intra-S-phase checkpoint response via an interaction with Rad53 in Saccharomyces cerevisiae. The Dbf4 protein has three highly conserved motifs, called the N, M and C motifs. In view of the fact that a comprehensive analysis of the roles of the three motifs in the initiation of DNA replication and checkpoint response was not previously available, this study was, therefore, conducted. The objectives of the study were: (1) to assess the function of the three conserved motifs, with respect to their essentiality for cell viability, (2) to determine their roles in mediating interactions with other proteins (i.e. Cdc7, Orc2, Mcm2) involved in the initiation of DNA replication and with Rad53 in the intra-S-phase checkpoint response, and (3) to obtain the three-dimensional structure of the Dbf4 N-motif by X-ray crystallography.
The Dbf4 N-motif was found to be nonessential for cell viability, mediates the interaction between Dbf4 and Rad53, and as well as the interaction with Orc2. A mutant lacking the N-motif (dbf4N), was found to have a growth defect and was hypersensitive to the genotoxic agents: hydroxyurea (HU) and methyl methane sulfonate (MMS), suggesting that a disruption in the intra-S-phase checkpoint occurred because of an abrogated Dbf4-Rad53 interaction. Double point mutation of two threonine residues of the N-motif (threonines 171 and 175) to alanines also caused an abrogated Dbf4-Rad53 FHA1 domain interaction.
The Dbf4 M-motif was found to be essential for cell viability and mediates the interaction between Dbf4 and Mcm2. A single proline to leucine point mutation at amino acid residue 277 conferred resistance to HU and MMS and caused disrupted Dbf4-Mcm2 and Dbf4-Orc2 interaction, while Dbf4-Rad53 interaction was maintained. Thus, the alteration of the M-motif may facilitate the role of Dbf4 as a checkpoint target.
The Dbf4-C motif was also found to be essential for cell viability. Deletion and point mutations to the C-motif affected the interactions between Dbf4 and Rad53, Orc2, Mcm2 and also with Mcm4.
Attempts were also made to obtain the three-dimensional structure of Dbf4, using X-ray crystallography methods.
The work presented here represents a thorough functional analysis of the three conserved domains of Dbf4 in Saccharomyces cerevisiae. These results can be used as a baseline for further research involving higher eukaryotic organisims, including humans. This is particularly of relevance in light of recent evidence demonstrating an overexpression of the human Dbf4 orthologue overexpression as a cancer phenotype in human cancer cells.
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