Journal of Biotechnology & Biomaterials
Open Access

Abstract

Human Polycystic Kidney Disease-1 (pkd1) gene promoter possesses various transcription factors in its vicinity which regulate its activity. The regulatory processes thus, layout the role of various transcriptional elements associated with the pkd1 gene in understanding the Autosomal Dominant Polycystic Kidney Disease (ADPKD) pathophysiology. These transcription factors either coalesce through coordinated interaction or work independently of each other hence maintaining pkd1 gene expression. There are substantial numbers of transcription factors which still remain to be identified. Identification of human pkd1 gene mutations within 5â�� untranslated region together with haplo-insufficiency has shifted the focus towards understanding gene regulatory pathways at pkd1 gene locus. As a result, 3.3-kb, -2.0-kb, -0.95 and -0.2-kb 5â�� proximal regions of the human pkd1 gene promoter were analyzed to posses Hepatocyte Nuclear Factor-1 (HNF-1�²) transcription factor binding site. The results show that HNF-1�² co-localized with P300/PCAF in nucleus and up-regulates pkd1 gene regulation. Deletion construct analysis revealed presence of HNF-1�² consensus sequences in the -0.95-kb region and posses a functional binding site through histone acetyl-transferase (p300/PCAF). Localization studies also confirmed its subcellular location to the nucleus using GFP with DAPI stained nuclei expressed in HEK293T cells. RT-PCR showed that overexpression of HNF-1�² treatment of HEK293T cells increased the levels of endogenous pkd1 RNA and EMSA and chromatin immunoprecipitation showed the presence of a novel HNF-1�² binding site at the pkd1 proximal promoter other than the typical evolutionary binding site. Therefore, these transcriptional studies and various molecular cross-talks bring out the mode of understanding ADPKD targeting the mechanism associated with HNF-1�² up-regulation.

Biography

Email: varsha_sh@hotmail.com