Macromolecular interaction on a cAMP responsive region in the urokinase-type plasminogen activator gene : a role of protein phosphorylation

• Published in: Nucleic acids research Vol. 18; no. 8; pp. 1991 - 1999
• Main Authors: VONDER AHE, D, PEARSON, D, NAGAMINE, Y
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 25.04.1990
• Subjects: Base Sequence; Binding, Competitive; Biological and medical sciences; Cell Line; Cyclic AMP - metabolism; DNA - genetics; DNA - metabolism; DNA-Binding Proteins - metabolism; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation, Enzymologic; Genes; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Phosphorylation; Plasminogen Activators - genetics; Plasminogen Activators - metabolism; Repetitive Sequences, Nucleic Acid; Restriction Mapping; Transcription, Genetic; Urokinase-Type Plasminogen Activator - genetics; Urokinase-Type Plasminogen Activator - metabolism; Binding protein; Urokinase; Regulation(control); DNA; Cyclic AMP; Restriction map; Molecular complex; Gene expression
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/18.8.1991

We have studied the regulation of urokinase-type plasminogen activator gene expression by cAMP in LLC-PK1 cells. We found a cAMP responsive region 3.4 kb upstream of the transcription initiation site, which comprised three protein-binding domains designated A, B, and C. Domains A and B both contain a sequence, TGACG, homologous to a consensus cAMP response element (CRE; TGACGTCA). Effective cAMP-mediated induction was achieved when these two domains were linked with domain C, which by itself did not confer cAMP responsiveness to a heterologous promoter nor contained CRE-like sequence, suggesting a functional cooperation among these domains. Results of competition studies using gel retardation and DNase I footprinting assays suggest that there is a protein-protein interaction between a CRE binding protein and a domain C binding protein. In gel retardation assays, binding of a nuclear protein to domains A and B was strongly augmented by addition of the catalytic subunit of cAMP-dependent protein kinase, whereas the protein binding to domain C was slightly inhibited, suggesting that protein phosphorylation is involved in the regulation of protein-DNA interaction.