The 5' terminal oligopyrimidine tract of human elongation factor 1A-1 gene functions as a transcriptional initiator and produces a variable number of Us at the transcriptional level.
The human elongation factor 1A-1 (eEF1A-1) gene is a member of the 5' terminal oligopyrimidine tract (5' TOP) gene family, and the number of thymidines (Ts) at the 5' TOP of cDNAs corresponding to this gene is known to show variation. Here we determined the 5'-end sequences of 125 eEF1A-1 clones and the complete sequences of 19 eEF1A-1 clones from an oligo-capped cDNA library and showed that variation in the number of Ts is generated by an in vivo process, not by an in vitro artifact during the construction of the cDNA library. Moreover, using green fluorescent protein transgenic mice, we demonstrated that the variation in T number is probably generated during or after transcription. We also introduced various mutations in the mRNA start site of this gene, particularly in the T stretch at the 5' TOP, and examined the effects on the promoter activity. The results showed that at least three Ts must exist at the 5' TOP for the high transcriptional activity of the eEF1A-1 gene promoter. Many other housekeeping genes, including ribosomal protein genes, are also members of the 5' TOP gene family, and the 5' TOP sequence may be an important core-promoter element of these genes.
Shibui-Nihei
Akiko
A
Department of Preventive Medicine, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto, Nagano 390-8621, Japan.
Ohmori
Yoshihiro
Y
Yoshida
Kenichi
K
Imai
Jun-ichi
J
Oosuga
Ikurou
I
Iidaka
Makoto
M
Suzuki
Yutaka
Y
Mizushima-Sugano
Junko
J
Yoshitomo-Nakagawa
Kiyomi
K
Sugano
Sumio
S
D020121:5' Untranslated Regions / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009154:Mutation; D020648:Peptide Elongation Factor 1 / Q000235:genetics*; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000235:genetics; D011742:Pyrimidine Nucleotides / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000235:genetics; D014500:Uracil Nucleotides / Q000235:genetics*
D020121:5' Untranslated Regions; D018076:DNA, Complementary; D008164:Luminescent Proteins; D020648:Peptide Elongation Factor 1; D011742:Pyrimidine Nucleotides; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014500:Uracil Nucleotides; D049452:Green Fluorescent Proteins; D015500:Chloramphenicol O-Acetyltransferase