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Isolation of the promoter region of the PHKG1 gene
Author(s) -
Elliott Natalina Elizabeth,
Rice Nancy A
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a957-b
Subject(s) - gene isoform , biology , gene , transcriptional regulation , protein subunit , promoter , regulatory sequence , reporter gene , binding site , regulation of gene expression , gene expression , microbiology and biotechnology , genetics
Phosphorylase kinase (PhK) is a key regulatory enzyme in glycogenolysis. With two isoforms, muscle and liver, this hexadecameric enzyme is coded for by eight unlinked genes. The catalytic subunit, ã, is sterically regulated by the á and â subunits and by an endogenous molecule of calmodulin, the ä subunit. Because of the high coordination of the expression among the four subunits, ã does not occur freely under normal physiological conditions as this would be detrimental to the system. To better understand the transcriptional regulatory mechanisms underlying PhK expression in muscle, the promoter region of the muscle isoform of the ã subunit (PHKG1) was isolated. Bioinformatic analyses were used to determine the putative promoter region. Comparison of the putative human promoter region to the previously isolated rat promoter reveals 48% sequence identity. While sequence conservation is not high, key binding sites have been evolutionarily conserved. Including two YY‐1 sites, a Nkx25 and 2 E‐box muscle specific protein binding sites. The human PHKG1 promoter (2.4 kb) has been ligated into a luciferase reporter vector and activity in both liver cells (HepG2) and muscle cells (C2C12) was determined. Such data will provide important information regarding the transcriptional regulation of PhK. This work was supported by Nataional Institutes of Health Center for Research. Resource Grant P2121216481.