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
- Full Text:
- Date Issued: 2015
- 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
- Full Text:
- Date Issued: 2015
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
- Full Text:
- Date Issued: 2015
- 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
- Full Text:
- Date Issued: 2015
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