Promoter analysis of the human translation termination factor 1 gene.
The human translation termination factor 1 (ETF1) gene encodes a class-1 release factor, eRF1, which catalyses termination of protein synthesis at all three stop codons. In this report, we describe the functional organization of the 5'-region of the gene. Primer extension and ribonuclease protection mapping revealed three transcription start sites clustered within approximately 10 bp. DNase I-hypersensitive site analysis identified five hypersensitive sites, one of which was located downstream of the initiation start sites. We used transient expression assays to define the 5'-regulating regions and in vivo and in vitro footprinting analysis to identify potential cis-acting regulatory elements. A basal promoter, spanning nucleotides -210/+117, contained no TATA box but a putative initiator element (Inr) and multiple potential Sp1/Sp3 binding sites, and thus displayed some of the features of a housekeeping gene. An additional upstream promoter containing positive and negative regulatory elements also regulated ETF1 gene expression. Real-time quantitative RT-PCR analysis showed tissue-specific expression of ETF1 transcripts in mouse tissues. Our results are suggestive of a constitutive expression of the human ETF1 gene but with possible cell- and tissue-specific regulation.
Dubourg
Christèle
C
Département de Biochimie et Biologie Moléculaire, UMR 6061 "Génétique et Développement", Faculté de Médecine, CS 34317, 2 Avenue du Pr. Leon Bernard, 35043 Cedex, Rennes, France.
Toutain
Bertrand
B
Le Gall
Jean Yves
JY
Le Treut
André
A
Guenet
Lucienne
L
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015139:Blotting, Southern; D019556:COS Cells; D002522:Chlorocebus aethiops; D018983:DNA Footprinting; D019175:DNA Methylation; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010454:Peptide Termination Factors / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D014162:Transfection
C489002:ETF1 protein, human; C489003:Etf1 protein, mouse; D010454:Peptide Termination Factors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D008156:Luciferases