Activation of c-myc promoter P1 by immunoglobulin kappa gene enhancers in Burkitt lymphoma: functional characterization of the intron enhancer motifs kappaB, E box 1 and E box 2, and of the 3' enhancer motif PU.
Deregulated expression of the proto-oncogene c- myc in Burkitt lymphoma (BL) cells carrying a t(2;8) translocation is mediated by a synergistic interaction of the translocated immunoglobulin (Ig) kappa gene intron (kappaEi) and 3' (kappaE3') enhancers and characterized by a strong activation of the promoter P1. We have investigated the functional role of distinct kappa enhancer sequence motifs in P1 activation on both mini-chromosomes and reporter gene constructs. Stable and transient transfections of BL cells revealed critical roles of the kappaEi and kappaE3' elements kappaB and PU, respectively. Joint mutation of kappaB and PU completely abolished P1 activity, implying that an interaction of kappaB- and PU-binding factors is essential for the enhancer synergism. Mutation of the E box 1 and E box 2 motifs markedly decreased P1 activity in transient but not in stable transfection experiments. Co-expression of the NF-kappaB subunit p65(RelA) and Sp1, an essential factor for P1 transcription, in Drosophila melanogaster SL2 cells synergistically enhanced promoter activity. Our results support a model which proposes cross-talk between promoter and enhancer binding factors as the basic mechanism for kappa enhancer-mediated c- myc activation in BL cells.
Wittekindt
N E
NE
GSF-National Research Center for Environment and Health, Institute of Clinical Molecular Biology and Tumor Genetics, Marchioninistrasse 25, D-81377 Munich, Germany. wittekin@biochem.mpg.de
Hörtnagel
K
K
Geltinger
C
C
Polack
A
A
D000818:Animals; D001483:Base Sequence; D002051:Burkitt Lymphoma / Q000235:genetics* / Q000473:pathology; D002460:Cell Line; D002875:Chromosomes / Q000235:genetics; D004331:Drosophila melanogaster / Q000166:cytology; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D016259:Genes, myc / Q000235:genetics*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics*; D007438:Introns / Q000235:genetics; D009154:Mutation / Q000235:genetics; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D020218:Response Elements / Q000235:genetics*; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D051996:Transcription Factor RelA; D015533:Transcriptional Activation / Q000235:genetics*; D014162:Transfection; D014178:Translocation, Genetic / Q000235:genetics
D007145:Immunoglobulin kappa-Chains; D016328:NF-kappa B; D016329:Sp1 Transcription Factor; D051996:Transcription Factor RelA