A novel transcriptional initiator activity of the GABP factor binding ets sequence repeat from the murine cytochrome c oxidase Vb gene.
The murine cytochrome c oxidase (COX) subunit Vb mRNA contains heterogeneous 5' ends mapping to +1, +6, +12, +17-22, +24-29, and +32-35 positions of the gene. We have previously shown that initiation of RNA at the +1 position of the promoter depends upon a YY-1 (NF-E1) binding initiator motif. In this article we show that the GABP factor binding duplicated ets motif, GTTCCCGGAAG, at +16 to +26 position functions as an independent initiator for transcription of RNAs mapping to the +18-19 and +23-26 regions. The initiation region ets sequence repeat (ets-ets sequence) can drive the transcription of the CAT reporter gene. The upstream ets site of the ets-ets sequence exhibits a low affinity for binding to purified GABP factors, whereas the downstream site exhibits high affinity. S1 analyses of RNA from transfected COS cells demonstrate that the initiation region ets-ets sequence can accurately initiate transcription at the +18-19 and +24-25 regions. Transcriptional initiation at these two positions, but not at +1, +12, and +31-32 positions, show a selective dependence for intact downstream ets site and GABP alpha and beta factors as tested in an in vitro reconstituted system. The activities of both COX IV and COX Vb single site ets initiators are induced in vivo by coexpression with GABP alpha and beta cDNAs. These results provide evidence that the 5' heterogeneity of the COX Vb mRNA is largely due to independent transcription initiators at multiple initiator motifs that bind to various transcription factors.
Sucharov
C
C
Department of Animal Biology, University of Pennsylvania, Philadelphia 19104-6047, USA.
Basu
A
A
Carter
R S
RS
Avadhani
N G
NG
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002874:Chromosome Mapping; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics*; D051268:GA-Binding Protein Transcription Factor; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011485:Protein Binding / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D051268:GA-Binding Protein Transcription Factor; C495656:Gabpa protein, mouse; C495658:Gabpb1 protein, mouse; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger; D014157:Transcription Factors; D003576:Electron Transport Complex IV