Molecular cloning and characterization of the human phosducin-like protein (hPhLP) promoter

摘要

Phosducin-like protein (PhLP) is an inducible Gβγ binding protein which is hypothesized to be a ubiquitous G protein regulator. To elucidate the mechanisms regulation the expression of the human PhLp (hPhLP) gene, we have cloned and characterized its 5'-flanking region. Primer extension analysis identified a major transcription initiation site 172 bp upstream of the ATG start codon. Analysis of the 5'-flanking region revealed that, although it lacked a TATA box element, the hPhLP promoter did contain several consensus binding motifs including AP4, CCAAT, CREB, NF-κB, SPl and E2F. Transient transfection analyses using a series of 5'-flanking deletion/luciferase reporter gene constructs identified a 25 bp sequence (-80 to -55 bp) that is necessary for basal level transcription of the hPhLP gene in all the cell lines investigated. Interestingly. Dependent upon the cell line, distinct transcription factors bind to this region suggesting that basal level hPhLP gene transcription may be regulated in a tissue-specific manner.