Molecular cloning of a functional promoter of the human plakoglobin gene

摘要: Objective Plakoglobin (Pg) is the only cytoplasmic protein component common to both junctional complexes mediating cell-cell adhesion, adherens junctions and desmosomes. In these Pg appears act as a linker anchoring transmembrane proteins of cadherin superfamily actin cytoskeleton intermediate filament system respectively. Intercellular adhesion frequently disturbed in skin diseases carcinomas, enabling tumour progression metastasis. Whereas expression lost some thyroid tumours carcinoma cell lines, little information on gene regulation currently available owing lack promoter studies. Design methods We have cloned sequenced genomic DNA from human library that resulted 979 bp upstream published cDNA. The transcriptional start was mapped by rapid amplification cDNA ends. Methylation-specific PCR bisulfite-modified line applied probe methylation status promoter-associated CpG island. Reporter-gene constructs various fragments were transiently transfected lines their activities determined luciferase measurements. Results conclusions A 1 kb fragment harbouring functional characterized. sequence lacks canonical TATA box, but contains putative CCAAT boxes well binding sites for transcription factors, among them SP1 AP2, proximal start. Considerable activity found deletion analysis indicated 300 region 5'-untranslated mRNA represents minimal gene. As cells lacking endogenous contain methylated dinucleotides island located around site, it suggested epigenetic mechanisms such contribute dysregulated expression.