Chromosomal integration dependent induction of junB by growth factors requires multiple flanking evolutionarily conserved sequences.
The junB locus contains nine flanking evolutionarily conserved sequences (FECS) that share 72% to 91% sequence identity between human and mouse. These FECS encompass the same regions of flanking DNA necessary for maximal mitogenic induction of junB. Most of the cis elements reported to date that affect junB regulation also reside within FECS. These observations suggest that the persistence of FECS through evolution reflects a necessary role in junB transcriptional regulation. In this report, we identify specific regulatory cis elements within junB FECS II and III and provide a quantitative analysis of the contribution made by these sequences to junB induction. These cis elements include a Serum Response Element (SRE), two Ets sites previously unrecognized as contributing to junB expression, and two novel Ets-linked motifs (ELMs). In general, mutating any single element significantly impairs junB induction. Moreover, the same mutations alter the structure of junB 5' flanking DNA within chromatin. Collectively, these results suggest that multiple proteins bound within FECS confederate to form a functional promoter complex, the activity of which is dependent upon a specific chromatin architecture.
Phinney
D G
DG
Fox Chase Cancer Center, Philadelphia, Pennsylvania 19007, USA.
Tseng
S W
SW
Hall
B
B
Ryder
K
K
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002843:Chromatin / Q000737:chemistry; D002875:Chromosomes / Q000235:genetics; D017124:Conserved Sequence; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods; D004815:Epidermal Growth Factor / Q000494:pharmacology; D019143:Evolution, Molecular; D015972:Gene Expression Regulation, Neoplastic; D016762:Genes, fos; D006133:Growth Substances / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D019868:Kanamycin Kinase; D051379:Mice; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D016755:Proto-Oncogene Proteins c-jun / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D026362:Serum Response Factor
D002843:Chromatin; D004268:DNA-Binding Proteins; D006133:Growth Substances; D009687:Nuclear Proteins; D009841:Oligonucleotides; D010982:Platelet-Derived Growth Factor; D016755:Proto-Oncogene Proteins c-jun; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; D026362:Serum Response Factor; C050379:oncogene proteins v-ets; D004815:Epidermal Growth Factor; D017853:Phosphotransferases (Alcohol Group Acceptor); D019868:Kanamycin Kinase