CHARACTERIZATION OF THE JOINING CHAIN (J-CHAIN) PROMOTER

A 300 b.p. promoter from the mouse joining chain (J-chain) gene was studied with regard to functional activity and protein/DNA interactions. The promoter only stimulated expression of a chloramphenicol-acetyl-transferase (CAT) reporter gene when an enhancer was present in the construct, regardless of whether the construct was transfected into cell lines that did or did not express an endogenous J-chain. Furthermore, deletion mutants lacking the 5' portion of the promoter were transcribed at a higher rate than the intact promoter in both J-chain positive and J-chain negative B-cell lines but not in untransformed B lymphocytes stimulated by lipopolysaccharide, indicating the presence of a negative control element in the 5' portion of the J-chain promoter active in tumour cells only. The octamer element in the J-chain promoter was found to bind Oct proteins, albeit with a low affinity. The penta-deca (p.d.) element in the J-chain promoter bound proteins in extracts from untransformed B cells but not in the tested cell lines. The protein binding to the J-chain p.d. element did not compete efficiently with a p.d. element from the SP6 kappa promoter. A protein binding to the 5' portion of the J-chain was expressed in some cell lines but not in others; neither a negative nor a positive correlation to J-chain expression could be seen. It was concluded that the J-chain promoter is equivalent to a kappa promoter and that differentiation-specific J-chain expression is governed by distal, positive control elements located outside the analysed region.