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Functional analysis of P1, a model R2R3 MYB domain transcription factorHeine, George F. January 2006 (has links)
Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 140-155).
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An analysis of CIS and trans-acting factors controlling bovine papillomavirus type 1 early transcriptionHarrison, Stephen Mark January 1988 (has links)
No description available.
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Analysis of predictive power of binding affinity of PBM-derived sequencesMatereke, Lavious Tapiwa January 2015 (has links)
A transcription factor (TF) is a protein that binds to specific DNA sequences as part of the initiation stage of transcription. Various methods of finding these transcription factor binding sites (TFBS) have been developed. In vivo technologies analyze DNA binding regions known to have bound to a TF in a living cell. Most widely used in vivo methods at the moment are chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) and DNase I hypersensitive sites sequencing. In vitro methods derive TFBS based on experiments with TFs and DNA usually in artificial settings or computationally. An example is the Protein Binding Microarray which uses artificially constructed DNA sequences to determine the short sequences that are most likely to bind to a TF. The major drawback of this approach is that binding of TFs in vivo is also dependent on other factors such as chromatin accessibility and the presence of cofactors. Therefore TFBS derived from the PBM technique might not resemble the true DNA binding sequences. In this work, we use PBM data from the UniPROBE motif database, ChIP-seq data and DNase I hypersensitive sites data. Using the Spearman’s rank correlation and area under receiver operating characteristic curve, we compare the enrichment scores which the PBM approach assigns to its identified sequences and the frequency of these sequences in likely binding regions and the human genome as a whole. We also use central motif enrichment analysis (CentriMo) to compare the enrichment of UniPROBE motifs with in vivo derived motifs (from the JASPAR CORE database) in their respective TF ChIP-seq peak region. CentriMo is applied to 14 TF ChIP-seq peak regions from different cell lines. We aim to establish if there is a relationship between the occurrences of UniPROBE 8-mer patterns in likely binding regions and their enrichment score and how well the in vitro derived motifs match in vivo binding specificity. We did not come out with a particular trend showing failure of the PBM approach to predict in vivo binding specificity. Our results show Ets1, Hnf4a and Tcf3 show prediction failure by the PBM technique in terms of our Spearman’s rank correlation for ChIP-seq data and central motif enrichment analysis. However, the PBM technique also matched the in vivo binding specificities of FoxA2, Pou2f2 and Mafk. Failure of the PBM approach was found to be a result of variability in the TF’s binding specificity, the presence of cofactors, narrow binding specificity and the presence ubiquitous binding patterns.
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Characterisation of the human α2(I) procollagen promoter-binding proteinsCollins, Malcolm Robert January 1993 (has links)
In an attempt to elucidate the transcriptional mechanisms that regulate the expression of the human α2(I) procollagen gene, cis-acting DNA-elements within the proximal promoter were identified and their corresponding trans-acting factors characterised. The fibroblast cell lines used in this study had previously been transformed with either simian virus 40 (SVWI-38) or by γ-radiation (CT-1). The SVWI-38 fibroblasts do not produce any α2(I) collagen chains, whereas the CT-1 cell line produces normal type I collagen. Previous studies suggested that trans-acting factor(s) may be responsible for the inactivation of the α2(I) procollagen gene in SVWI-38 fibroblasts (Parker et. al. (1989) J. Biol. Chem 264, 7147-7152; Parker et. al. (1992) Nucleic Acids Res. 20, 5825-5830). In this study, the SVWI-38 proximal promoter (-350 to +54) was sequenced and shown to be normal, thereby ruling out any possibility that mutations within this region was responsible for inactivation of the gene.
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Expanding the Known DNA-binding Specificity of Homeodomains for Utility in Customizable Sequence-specific Nucleases: A DissertationChu, Stephanie W. 24 May 2013 (has links)
Homeodomains (HDs) are a large family of DNA-binding domains contained in transcription factors that are most notable for regulating body development and patterning in metazoans. HDs consist of three alpha helices preceded by an N- terminal arm, where the third helix (the recognition helix) and the N-terminal arm are responsible for defining DNA-binding specificity. Here we attempted to engineer the HDs by fully randomizing positions in the recognition helix to specify each of the 64 possible 3’ triplet sites (i.e. TAANNN). We recovered HD variants that preferentially recognize or are compatible with 44 of the possible sites, a dramatic increase from the previously observed range of specificities. Many of these HD variants contain combinations of novel specificity determinants that are uncommon or absent in extant HDs, where these determinants can be grafted into alternate HD backbones with an accompanying alteration in their specificity. The identified determinates expand our understanding of HD recognition, allowing for the creation of more explicit recognition models for this family. Additionally, we demonstrate that HDs can recognize a broader range of DNA sequences than anticipated, thus raising questions about the fitness barrier that restricts the evolution HD-DNA recognition in nature. Finally, these new HD variants have utility as DNA-binding domains to direct targeting of customizable sequence-specific nuclease as demonstrated by site-specific lesions created in zebrafish. Thus HDs can guide sequence-specific enzymatic function precisely and predictably within a complex genome when used in engineered artificial enzymes.
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Induction of apoptosis or cell cycle arrest by two human wildtype variants of the p53 proteinAzoulay, Eric. January 1999 (has links)
No description available.
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On the evolutionary origin of angiosperms : characterization of MADS-box floral homeotic gene homologues in Ephedra andina (Gnetales)Savard, Joël. January 2000 (has links)
No description available.
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The evolution of meiotic recombination in vertebrates: the case of snakesHoge, Carla R. January 2024 (has links)
Comparisons among model organisms make clear that, despite the fundamental importance of recombination in sexually-reproducing species, the mechanisms by which it is directed to the genome can vary markedly. Notably, in mice and humans, recombination almost exclusively occurs where the protein PRDM9 binds DNA. In such species, fine-scale recombination rates along the genome are rapidly evolving, as shifts in PRDM9 binding affinity remodel the landscape. In other species such as birds or canids, PRDM9 has been lost and recombination occurs preferentially at promoter-like features, leading to the conservation of recombination rates over large evolutionary distances. Increased recombination near promoters is also seen in human and mouse knockouts for PRDM9, indicating that this mechanism is normally out-competed by PRDM9 binding. The rapid evolution of complete orthologs of PRDM9 in non-mammalian vertebrates suggests that the protein may play a similar role in directing recombination outside of mammals.
In chapter 2 of this work, we test this hypothesis by focusing on the corn snake Pantherophis guttatus, a representative vertebrate species with a single, complete PRDM9 ortholog that is rapidly evolving. We improved the assembly and annotation of the corn snake reference genome and resequenced 24 unrelated corn snake samples to high coverage in order to infer historical recombination rates across the genome from patterns of linkage disequilibrium. We find evidence for elevated recombination around computationally predicted PRDM9 binding sites but, surprisingly, also near promoter features. To verify these findings, we resequenced two pedigrees, identified the PRDM9 alleles segregating in the families and called crossover events that occurred in the parents.
This analysis confirmed that crossover events overlap both PRDM9 binding sites and promoter features more than expected by chance. Thus, unlike in mammalian species that rely on PRDM9, in corn snakes there appears to be a mixed use of PRDM9 binding sites and promoter like features, and we find evidence that the relative importance of these features differs between macro- and microchomosomes. We hypothesize that the dual usage of these features reflects a tug of war between PRDM9 and promoter features, whose strength in snakes and possibly other vertebrates has been shifted by changes to a gene that reads the histone modifications made by PRDM9, and likely other genes. In chapter 3, I discuss how follow-up experiments based on these observations could help answer long-standing questions related to the conditions under which PRDM9-directed recombination localization is favorable. Beyond the specific results, this work illustrates how the study of non-model organisms can inform our understanding of basic genetic mechanisms.
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Determining the Molecular Function of a Translesion DNA Synthesis ComplexTetenych, Andriana January 2022 (has links)
Translesion DNA Synthesis (TLS) is a mechanism that promotes DNA damage tolerance during DNA replication using an error-prone DNA polymerase complex. The complex is comprised of the ImuA, ImuB, and ImuC proteins that are found in approximately one-third of bacteria, including high priority antimicrobial resistant pathogens, such as Pseudomonas aeruginosa. Previous in vivo studies have shown that TLS increases beneficial bacterial mutations as the error-prone DNA polymerase, ImuC, lacks proof-reading activity. However, how ImuA and ImuB proteins contribute to the polymerase mechanism is unknown. Thus, the goal of this study is to characterize the TLS proteins in vitro to determine how ImuA and ImuB associate with ImuC to promote error- prone replication.
ImuA and ImuBNΔ34 were successfully purified for biochemical characterization from the homolog Myxococcus xanthus. Using size-exclusion chromatography coupled to multi-angle light scattering, both ImuA and ImuBNΔ34 are trimers in solution. Each protein also binds DNA independently as assessed by fluorescence polarization. Interestingly, both proteins bind ssDNA and a 3’ overhang substrate mimicking the DNA replication intermediate with the highest affinity. DNA binding assays further confirm these proteins can form a DNA-ImuA-ImuBNΔ34 complex. Using bacterial two-hybrid assays, the ImuA- ImuB interaction occurs in the C-terminal region of both proteins. Overall, these results suggest that ImuA and ImuB may recruit and stabilize ImuC on DNA for replication past damaged DNA, providing the first insights into the ImuA and ImuB molecular mechanism. / Thesis / Master of Science (MSc)
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Characterization of the DNA Binding Properties of CST (CTC1-STN1-TEN1) And Their Importance for CST Function in Telomeric as well as Genome-wide ReplicationBhattacharjee, Anukana, M.S. 05 December 2017 (has links)
No description available.
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