Biochemical and Biophysical Research Communications, Vol.329, No.2, 755-764, 2005
Identification and in silico characterization of a novel gene: TPA induced trans-membrane protein
12-O-Tetradecanoylphorbol-13-acetate (TPA) is a potent tumor promoter with wide ranging, diverse, and sometimes opposite cellular effects. Using oligonucleotide microarray analysis, we have identified a novel gene that is upregulated following treatment with TPA in the pancreatic cancer cell line CD18. Real-time PCR validated the microarray results in CD18 and HeLa cells, and showed that upregulation of the gene is time- and concentration-dependent. In silico analysis showed the gene product to be a single-pass transmembrane protein of 217 residues that is localized to the endoplasmic reticulum, thus the name TPA induced trans-membrane protein (TTMP). A luciferase reporter assay demonstrated that upregulation of TTMP by TPA is triggered at the promoter level. Published by Elsevier Inc.
Keywords:12-O-tetradecanoylphorbol-13-acetate;protein kinase C;TTMP;novel protein;microarray;real-time PCR;in silico analysis;luciferase assay