I= SOCIO – ECONOMIC CONSEQUENCES OF RETURN MIGRATION AND THE ADJUSTMENT PROBLEMS IN PUNJAB Title of Thesis


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Title of Thesis

AML AS TRANSCRIPTIONAL REGULATOR AND OSTEOBLAST DIFFERENTIATION MEDIATOR

Author(s)

Amjad Javed

Institute/University/Department Details

University of the Punjab, Lahore / Zoology

Session

2000

Subject

Zoology

Number of Pages

320

 

Keywords (Extracted from title, table of contents and abstract of thesis)

AML, rat osteocalcin, vitamin D response element (VDRE), Osteoblast, Bone resorption, DNA

 

Abstract

Three AML motifs are strategically positioned in the bone-specific rat osteocalcin (OC) promoter. Sites A and B flank the vitamin D response element (VDRE) in the distal promoter and sites Band C define the borders of a positioned nucleosome in the proximal promoter. The functional significance of each AML element was addressed by mutating individual or multiple AML sites within the context of the native rat (-1.1 kb) OC promoter-CAT. Reporter gene activity was assayed following transient transfection and after stable genomic integration in ROS 17/2.8 osteoblastic cell lines. It has been shown that all three AML sites are required for maximal basal expression of the rOC promoter. Furthermore, the distal sites A and B can functionally compensate for a mutation at the proximal site C and paired mutations involving site A (mAB or mAC) result in a loss of activity nearly equivalent to mutation of all three sites (mABC). Strikingly, mutation of the three AML sites leads to abrogation of responsiveness to vitamin D, glucocorticoids and TGF-B. We also find mutation of the three AML sites results in loss of the DNase hypersensitive sites at the VDRE and over the proximal tissue-specific basal promoter. These findings strongly support a multifunctional role for AML factors in regulating gene expression, not only as a simple transactivator, but also by facilitating modification in promoter architecture and chromatin organization.

 

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    Table of Contents  

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1 1 INTRODUCTION 1

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2 2 MATERIALS AND METHODS 140

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3 3 ABSTRACT 193

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4 4 REFERENCES 275

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5 5 BIBLIOGRAPHY

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