Thesis (Ph.D. in Biochemistry and Molecular Genetics) -- University of Colorado, 2005. / Typescript. Includes bibliographical references (leaves 84-97). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

Thesis (Ph.D.)--Hong Kong University of Science and Technology, 2006. / Includes bibliographical references (leaves 148-167). Also available in electronic version.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 102-120).

by Brian C. Freeman. / Typescript. / Thesis (M.S.)--Florida State University, 1991. / Includes bibliographical references.

Abstract The human and mouse genes POLE1 and Pole1 for the catalytic subunit of DNA polymerase ε contain 51 and 49 exons, respectively, and the human gene POLE2 for the B-subunit contains 19 exons. The human POLE1 encodes three alternatively spliced mRNAs differing in their 5'-terminal sequence and in the N-termini of the predicted proteins. The promoters for the major human transcript and the mouse Pole1 are G+C rich, TATA-less and contain putative cis-acting elements typical of both S phase upregulated and serum responsive promoters. Interestingly, the three human alternative transcripts are expressed from three promoters, and other structural features of POLE1 suggest that regulation of its expression is complicated. The amino acid sequence of the catalytic subunit deduced from the mouse cDNA shows remarkable evolutionary conservation in the DNA polymerase ε family. Interestingly, several conserved elements involved in template-primer binding differ from those of other class B DNA polymerases. This is likely to reflect a distinctive function of the enzyme. The mouse Pole1 was localized to chromosome 5 region E3-E5. The expression of the human POLE2 encoding the B-subunit of DNA polymerase ε is dependent on cell proliferation in a late serum-responsive manner. This is typical for DNA replication-related proteins. The promoter, which utilizes multiple transcriptional initiation sites, is G+C rich and lacks a TATA-box. A 75 bp core promoter region is located within exon 1 and contains an Sp1 element as a critical determinant of the promoter activity. Two overlapping E2F elements adjacent to the Sp1 element are essential for full promoter activity and serum response. Immediately downstream from the core promoter region reside binding sites for E2F1 and NF-1. POLE2 seems to be regulated by two E2F-pocket protein complexes, one associated with Sp1 and the other with NF-1. The complete cDNA of the human DNA topoisomerase IIβ binding protein (TopBP1) reveals a 170 kDa protein that contains eight BRCT-domains and shows homology to S. cerevisiae Dpb11 and S. pombe Cut5/Rad4. The protein interacts physically with human DNA polymerase ε as shown by co-immunoprecipitation. A peptide containing the 6th BRCT-domain and an antibody against this peptide inhibit DNA replication in isolated nuclei, indicating that the protein is required for DNA replication. The expression of TopBP1 is proliferation-dependent in a manner that is typical for replication proteins. The gene encoding TopBP1 was localized to chromosome 3q21-q23.

L'entrée en sénescence, considérée comme un arrêt irréversible du cycle, se caractérise par une modification de l'organisation de la chromatine formant de véritables foyers d' hétérochromatine spécifiques (SAHF) coordonnée à une modification d'expression génique et à un déclin progressif de la compétence à répliquer le génome. Ainsi, au cours de ma thèse, j'ai voulu comprendre en quoi ces changements d'organisation du génome pouvaient influer sur la distribution et l'activation des origines d e réplication lors de l'entrée en sénescence réplicative ou déclenchée de façon prématurée par l'inhibition d'un modulateur de chromatine, la protéine à activité Histone AcétylTransférase p300. Pour étudier ces régulations, j'ai utilisé le peignage moléculaire d'ADN réplicatif qui permet de suivre les fourches de réplication et d'évaluer la distribution moyenne des origines. De plus, à l'aide de la purification de brins naissants aux origines de réplication couplée à un séquençage haut débit, nous avons cartographié la position de ces origines sur l'ensemble du génome humain et étudier un ensemble de facteurs pouvant intervenir dans ce déterminisme. Grâce à cette étude, nous avons pu suivre finement les modifications d'activité des origines associées à l'entrée en sénescence. De plus, afin de mieux comprendre les mécanismes d'activation des origines de réplication, nous avons étudié en collaboration avec l'équipe du Dr Fisher, le rôle de Cdk1 et de Cdk2 dans l'activation des origines dans le modèle Xénope. / Senescence entry, considered as an irreversible cell cycle arrest, is characterized by modifications of chromatin organization forming specific heterochromatin foci (SAHF) coordinated to modification of gene expression and the progressive loss of capacity to replicate the genome. During my PhD, we investigated whether these changes in genome organization might induce modifications in the distribution and the activity of replication origins during replicative senescence entry and in prematurely induced senescence by inhibition of a chromatin modulator, the Histone AcetylTransferase p300. To study these regulations, we used the replicating DNA combing allowing to follow the progression of replication forks and to evaluate the mean distribution of origins. By using the nascent strand purification assay coupled to deep sequencing, we mapped the position of replication origins in the whole human genome and studied some factors which could be involve d with this determinism. Thanks to this study, we followed finely the modifications of activity of replication origins associated to senescence entry. Moreover, in order to better understand the mechanisms of activation of origins, we studied in collaboration with Dr Fisher's team, the role of Cdk1 and Cdk2, in the activity of replication origins in the Xenopus model.

Previous research shows that bodily experiences can unconsciously influence perception, judgment, and behavior. However, inconsistency among recent findings in the embodied cognition literature suggests a need for theoretical boundary conditions. While research appears to assume that embodied effects are necessarily implicit (Schnall, 2017), the extant literature has not directly manipulated the role that conscious awareness of bodily states plays in embodied cognition. Dual process theories of social cognition assert that information processing falls along a continuum, from processing that is relatively automatic, effortless, and experiential, to processing that is relatively deliberate, controlled, and rational. Importantly, information processed along the dimensions of this continuum can lead to different outcomes. Thus, if the body influences social cognition in a more implicit manner, experimentally manipulating conscious awareness of a bodily state may lend further insight into when embodiment is attenuated. Six studies tested this possibility in the case of the demonstrated effect of weight sensations on judgments of an abstract idea’s importance (e.g., Ackerman, Nocera, & Bargh, 2010; Jostmann, Lakens, Schubert, 2009). Studies 1 and 2 revealed a curvilinear relationship between increased clipboard weight and ratings of importance such that participants rated a topic as more important when holding a moderately heavy, compared to light, clipboard; however, the importance ratings decreased when the clipboard was very heavy. This curvilinear relationship was not caused by a negative evaluation of the topic or the activation of a different metaphor (burden). In Study 3, ratings of importance increased with a moderately heavy clipboard compared to a light clipboard, but this difference was eliminated by explicitly drawing perceiver's attention to the weight of the clipboard. Study 4 extended the model and showed that even a very heavy clipboard can act as an embodiment of importance when participants are prevented from deliberately processing the weight of the clipboard via a cognitive load manipulation. Study 5 provided limited evidence establishing the role of cognitive motivation in embodiment as measured by need for cognition. However, experimentally manipulating cognitive motivation in Study 6 showed that individuals with higher cognitive motivation were more likely to show the embodied effect when the heft of the clipboard was subtle (i.e., holding a moderately heavy clipboard) whereas those with lower cognitive motivation were more likely to show the embodied effect when the heft of the clipboard was blatant (i.e., holding a very heavy clipboard). Collectively, these studies suggest that embodiment is subject to dual-processes whereby if something in the context draws conscious attention to a stimuli that activates an embodied metaphor, perceivers will no longer use their body as a source of information when processing the stimuli.

No description available.

Inconsistency among findings in the embodied cognition literature suggests a need for theoretical boundary conditions. The current research proposes that conscious attention of a bodily state can moderate its influence on social judgment. Three studies tested this possibility in the case of the demonstrated effect of weight sensations on judgments of an abstract idea's importance. Studies 1 and 2 showed that participants rated a topic as more important when holding a moderately heavy, compared with light, clipboard. However, when the clipboard was very heavy, participants rated the survey topic as less important compared with when the clipboard was moderately heavy. The differences in importance ratings were not caused by derogation of the topic or the activation of a different metaphor. In Study 3, the importance rating difference between light and moderately heavy clipboards was eliminated by explicitly drawing perceiver's attention to the clipboard's weight. Implications and future directions are discussed.

No description available.