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Up regulation of caveolin-1 during H2O2 induced oxidative stress

Dasari, Arvind (2005) Up regulation of caveolin-1 during H2O2 induced oxidative stress. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

Caveolae ("little caves") are 50 -100 nm invaginations seen on the plasma membrane of most cells. Caveolin-1, the marker protein of caveolae in most tissues, is the structural and functional unit of caveolae. Previous studies have demonstrated the tumor suppressor capability of caveolin-1. Our lab has already shown that expression of caveolin-1 induces premature senescence in cells, possibly including those with malignant potential. We proposed that this might explain the tumor suppressor function of caveolin-1. We have further shown that oxidative stress induces premature senescence through up regulation of caveolin-1. To further elucidate the molecular mechanisms underlying this process, we used H2O2 as a model to generate stress induced premature senescence (SIPS) and examined the response of caveolin-1 promoter under these conditions. Constructs with serially truncated segments of the mouse caveolin-1 gene promoter linked to a luciferase reporter gene were made and luciferase assays were carried out. These experiments demonstrated that the critical regions lay in two segments: -222/-372 and -91/-150 of the promoter region. A consensus Sp1 binding sequence was identified within each deleted segment. Gel shift analysis of protein binding from nuclear extracts to these caveolin promoter DNA sequences confirmed that transcription factors were binding to the Sp1 consensus elements as part of the transcriptional response to H2O2 induced senescence. Further deletion mutagenesis of the individual Sp1 consensus sites confirmed the identity of the transcription factor to be Sp1. These findings suggest that Sp1 mediates oxidant induced up regulation of caveolin-1 expression. In subsequent experiments, we examined the effect of inhibitors of p38 Mitogen activated protein kinase pathway on the levels of caveolin-1 expression during SIPS by western blot and luciferase assay. We found that this pathway plays a direct role in the up regulation of caveolin-1 during SIPS, possibly through modification of Sp1 to increase its activity.Public Health Importance: Cancer is a leading public health concern. Cav-1 has been shown to be a tumor suppressor gene involved in a large number of human tumors. Induction of premature senescence in cells with malignant potential is thought to be a vital tumor suppressor function. Our study aims to define the tumor suppressor capacity of Cav-1 by elucidating the pathway by which it induces premature senescence.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Dasari, Arvindarvind_dasari@hotmail.com
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee MemberFeingold, Eleanoreleanor.feingold@hgen.pitt.eduFEINGOLD
Committee MemberGalbiati, Ferrucciofeg5@pitt.eduFEG5
Committee MemberFerrell, Robert Erferrell@mail.hgen.pitt.eduRFERRELL
Date: 8 July 2005
Date Type: Completion
Defense Date: 2 June 2005
Approval Date: 8 July 2005
Submission Date: 9 June 2005
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Institution: University of Pittsburgh
Schools and Programs: Graduate School of Public Health > Human Genetics
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: Caveolin-1; caveolin-2; caveolin-3; EMSA; free radicals; oxidative stress; oxidative stress; transcription factor; p38MAPK; Sp1; stress induced premature senescence
Other ID: http://etd.library.pitt.edu/ETD/available/etd-06092005-234926/, etd-06092005-234926
Date Deposited: 10 Nov 2011 19:46
Last Modified: 15 Nov 2016 13:44
URI: http://d-scholarship.pitt.edu/id/eprint/8050

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