Identification of a functional promoter element in the 5'-flanking region of the rat cMG1/TIS11b gene.
The cMG1 gene was originally identified as a mitogen-stimulated primary response gene. However, in contrast to genes such as c-fos and TIS11, cMG1 is also expressed at significant levels before and after the transient elevation induced by agonists. We have sequenced a 1.3 kb rat genomic cMG1 clone, which includes 931 bp upstream of the transcription start site identified by primer-extension analysis. A 1033 bp fragment, including this 5'-flanking sequence, directed the expression of the reporter gene chloramphenicol acetyltransferase (CAT) in transfected NIH-3T3 cells. Progressive 5'-to-3' deletion indicated that an element located between -138 and -114 was responsible for most of this basal CAT expression. DNA mobility-shift assays showed that the sequence between -143 and -105 contained binding sites for cellular proteins, the principal complexes involving nucleotides between -119 and -105. We conclude that these complexes may represent the transcription factor-DNA element interactions that determine basal cMG1 expression.
Corps
A N
AN
Department of Cellular Physiology, Babraham Institute, Cambridge, U.K.
Pascall
J C
JC
Hadfield
K M
KM
Brown
K D
KD
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D051817:Butyrate Response Factor 1; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry; D004268:DNA-Binding Proteins; D017353:Gene Deletion; D015870:Gene Expression; D017930:Genes, Reporter; D017874:Immediate-Early Proteins; D051379:Mice; D008969:Molecular Sequence Data; D016716:PC12 Cells; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017422:Sequence Analysis, DNA; D014162:Transfection; D051816:Tristetraprolin
D051817:Butyrate Response Factor 1; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D011506:Proteins; D012333:RNA, Messenger; D051816:Tristetraprolin; C497553:Zfp36 protein, mouse; C497554:Zfp36 protein, rat; C497555:Zfp36l1 protein, rat; D015500:Chloramphenicol O-Acetyltransferase