The transcriptional regulation of the prothrombin gene expression in neuroblastomas was investigated because of the interest in non-hepatic thrombin expression and function in the nervous system. The data indicated that the murine prothrombin gene was distinctively transcribed in proliferating murine N2a cells and that the transcripts were decreased during the differentiation of N2a cells. The gene transcription in proliferating N2a cells was due to the C-I nuclear complex formation in the promoter region, -248/-140. Mutation analyses indicated that nucleotides from -237 to -231 are the core C-I binding site while the longer sequence -248/-140 is needed for the C-I binding. The C-I binding to the promoter -248/-140 could be inhibited by the presence of competitor probe -187/-166, and the mutation in nucleotides from -186 to -179 significantly diminished not only the formation of C-I binding in the promoter region but also the promoter activity in proliferating neuroblastoma cells. Cyclic AMP response element (CRE) modulator, CREM, appeared to selectively bind to the sequence encompassing -186/-179. Taken together, the results indicate that the prothrombin gene transcription in proliferating N2a cells was critically dependent on the cooperative interaction between the factor(s) binding to the C-1 cis-acting element (-237/-231) and the putative CRE site (-186/-179) in the prothrombin promoter, and that the lack of prothrombin expression that coincided with nerve differentiation was mainly due to the lack of C-I complex formation in the promoter.
Copyright 2004 Elsevier B.V.