Human FN1 is regulated by the heat-shock response
- Authors: Dhanani, Karim Colin Hassan
- Date: 2015
- Subjects: Uncatalogued
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/193487 , vital:45336
- Description: Heat shock protein 90 (Hsp90) and heat shock factors (HSFs) are known to be involved in the epigenetic regulation of several fundamental oncogenic genes. Fibronectin (FN) is an extracellular matrix (ECM) glycoprotein which plays key roles in cell adhesion and migration. Hsp90 binds directly to FN and Hsp90 inhibition has been shown to regulate FN protein levels and matrix formation. Where inhibition of Hsp90 with a C-terminal inhibitor (novobiocin) induced the loss of FN matrix, treatment with an N-terminal inhibitor (geldanamycin) increased FN matrix levels. GA treatment induced a strong dose and time dependent increase in FN1 promoter activity and increased total FN mRNA respectively. By contrast, NOV showed no increase in the promoter activity and no change in the expression of FN mRNA. As GA is known to induce the stress response, we investigated the relationship between the cell stress machinery and the transcriptional regulation of FN. Three putative heat shock elements (HSEs) were identified in the FN1 promoter. The loss of two of the three identified putative HSEs resulted in a loss in the basal transcriptional activity of the FN1 promoter in our reporter model. This was in addition to the loss of the induction of transcriptional activity with GA treatment observed with the full-length promoter. Binding of HSF1 to one of the putative HSEs, which was identified as potentially functional from the truncation analysis, was confirmed using ChIP. The occupancy of this HSE by HSF1 was shown to increase with GA treatment. These data support the hypothesis that FN1 is a functional HSF1 target gene. The 5' promoter regions of seven additional ECM protein encoding genes were analysed and mRNA levels were detected by quantitative RT-PCR upon treatment with GA. Collagen 4 _2 and laminin _3 mRNA were found to increase in the presence of GA, whereas collagen 4 _3 and osteopontin showed no change. Similarly to FN1, these data indicate that a subset of ECM genes may be under the regulation of the HSF1 mediated heat-shock response. This may have implications for our understanding of ECM dynamics in cancer, where the clinical application of Hsp90 inhibitors is intended. Additionally, our data provide a poten- tial underpinning for the role of the HSF1 mediated heat-shock response in several fibrotic and metabolic stress related pathologies. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2015
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If I Stay Right Here
- Authors: Ngamlana, Chwayita
- Date: 2015
- Subjects: Uncatalogued
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/193498 , vital:45337
- Description: My novella concerns the dynamics of a relationship between two girls. It shows the heterosexual‐like nature of the relationship rather than dwelling on the pressures on lesbians from society as a whole. At its core is the raw emotion and passion of the relationship, which is at the same time toxic, destructive and volatile because of their class differences and other insecurities. The work is influenced by the grit, openness, and innovation of several contemporary writers. , Thesis (MA) -- Faculty of Humanities, Institute for the Study of English in Africa, 2015
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The reform of world order?: BRICS in an Interpolar world
- Authors: Phyllis, Yvonne
- Date: 2015
- Subjects: Uncatalogued
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/193520 , vital:45339
- Description: International relations has recently seen new developments which are unpredictable and in their infant stage. The causes of these developments are plentiful, the consequences stemming from waning unipolarity to the emergence of new fora creating their own spaces. Whatever the case, international relations is not as it was in the period just after the Cold War. As a result, the terms of global governance established after 1990 have come under serious scrutiny. This transition has the makings of a new world order, an interpolar world order. This study asserts that we no longer live in a unipolar world, nor do we live in a world which can only be described as multipolar. It makes the argument that although multipolarity is a crucial element of the world, it only offers a partial description of today’s order. The study asserts that it is interpolarity which is closest in accounting for today’s world. Not only does it describe the world as multipolar, it also describes it as interdependent. To this end, the study provides a detailed account of what is meant by an interpolar world order and how differently it explains international events. It also provides an account of factors which can develop in an interpolar world. One of these includes allowing room for emerging powers to create their own spaces in efforts of avoiding co-option while continuing to realise the importance of operating within a context of continuity. This means that emerging countries create their own spaces but they also realise the importance of working with already established regimes such as the G20. BRICS (Brazil, Russia, India, China and South Africa) is used as a case study to indicate one of the new developments afforded by an interpolar order-the emergence of new fora. The study asserts that although BRICS is rife with internal dissimilarities, it has indicated some degree of political will in one aspect; that is; advocating for the reform of the terms of global governance, advocating for more representative forms of global governance. The study thus explores factors of a world whose contours are gradually changing but which are unpredictable and in flux. , Thesis (MA) -- Faculty of Humanities, Political and International Studies, 2015
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The role of Stress Inducible Protein 1 (STI1) in the regulation of actin dynamics
- Authors: Beckley, Samantha Joy
- Date: 2015
- Subjects: Heat shock proteins , Molecular chaperones , Actin , Microfilament proteins , Cell migration , Adenosine triphosphatase , Metastasis
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/193941 , vital:45409
- Description: Stress-inducible protein 1 (STI1) otherwise known as Hop (Hsp70/Hsp90 organising protein) is a highly conserved abundant co-chaperone of the Hsp70 and Hsp90 chaperones. STI1 acts as an adapter protein, where it regulates the transfer of protein substrates from Hsp70 to Hsp90 during the assembly of a number of chaperone-client protein complexes. The role of STI1 associating independently with non-chaperone proteins has become increasingly prominent. Recent data from colocalisation and co-sedimentation analyses in our laboratory suggested a direct interaction between STI1 and the cytoskeletal protein, actin. However, there was a lack of information on the motifs which mediated this interaction, as well as the exact role of STI1 in the regulation of cytoskeletal dynamics. Two putative actin binding motifs, DAYKKK (within the TPR2A domain) and a polyproline region (after the DP1 domain), were identified in mammalian STI1. Our data from in vitro interaction studies including surface plasmon resonance and high speed co-sedimentation assays suggested that both TPR1 and TPR2AB were required for the STI1-actin interaction, and peptides corresponding to either the DAYKKK or the polyproline motif, alone or in combination, could not block the STI1-actin interaction. Full length mSTI1 was shown to have ATPase activity and when combined with actin an increase in ATPase activity was seen. Ex vivo studies using STI1 knockdown shRNA HEK293T cells and non-targeting control shRNA HEK293T cells showed a change of F-actin morphology as well as reduction in levels of actin-binding proteins profilin, cofilin and tubulin in the STI1 knockdown cells. These data extend our understanding of the role of STI1 in regulating actin dynamics and may have implications for cell migration. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2015
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