TY - JOUR
T1 - Activation of a dual adenovirus promoter containing nonconsensus TATA motifs in Schizosaccharomyces pombe
T2 - Role of TATA sequences in the efficiency of transcription
AU - Swaminathan, Sathyamangalam
AU - Malhotra, Pawan
AU - Manohar, Chitra F.
AU - Dhar, Ravi
AU - Thimmapaya, Bayar
N1 - Funding Information:
This work was supported by Public Health Service grants AI 20156 and AI18029 from the National Institutes of Health. We thank N.Jones for introducing us to the S.pombe system, M.Furtado and Urmila Shinde for help in certain experiments, P.Liebermann, A.J.Berk, A.Hoffman and R.G.Roeder for the TBP expression plasmids, Jo Ann Wise for providing S.pombe growth protocols and John Benson for critical reading of the manuscript.
PY - 1993/6/11
Y1 - 1993/6/11
N2 - The role of TATA elements in the expression of a mammalian promoter was investigated in the fission yeast Schizosaccharomyces pombe, by studying the human adenovirus E2-early promoter. This is a unique dual promoter with two nonconsensus TATA elements directing transcription from two cap sites, +1 and -26. A sequence TTAAGA provides the TATA box function for the +1 promoter, whereas a sequence TαTT, with a closer resemblance to the consensus (TAT-AA/TA) provides this function for the -26 promoter. Yet, in human cells, the +1 promoter is transcribed about 20 fold more efficiently than the -26 promoter. We found that both promoters are transcribed faithfully in S.pombe with start sites identical or close to those found in human cells. Surprisingly, the relative ratio of expression for the +1 and -26 promoters was exactly reversed in S.pombe cells. This reversal appeared to be due to the relatively weak binding of S.pombe TATA binding protein to the TTAAGA motif, rather than to its rate of dissociation. Furthermore, we show that in S.pombe, promoter expression correlates well with the nucleotide sequence of the TATA element rather than the context in which it is placed. By contrast, it is the context of the TATA element, rather than its nucleotide sequence that appears to be critical for promoter expression in human cells. Our data suggest the existence of one or more additional factors in human cells that permit the utilization of nonconsensus TATA elements. S.pombe appears to lack these factors.
AB - The role of TATA elements in the expression of a mammalian promoter was investigated in the fission yeast Schizosaccharomyces pombe, by studying the human adenovirus E2-early promoter. This is a unique dual promoter with two nonconsensus TATA elements directing transcription from two cap sites, +1 and -26. A sequence TTAAGA provides the TATA box function for the +1 promoter, whereas a sequence TαTT, with a closer resemblance to the consensus (TAT-AA/TA) provides this function for the -26 promoter. Yet, in human cells, the +1 promoter is transcribed about 20 fold more efficiently than the -26 promoter. We found that both promoters are transcribed faithfully in S.pombe with start sites identical or close to those found in human cells. Surprisingly, the relative ratio of expression for the +1 and -26 promoters was exactly reversed in S.pombe cells. This reversal appeared to be due to the relatively weak binding of S.pombe TATA binding protein to the TTAAGA motif, rather than to its rate of dissociation. Furthermore, we show that in S.pombe, promoter expression correlates well with the nucleotide sequence of the TATA element rather than the context in which it is placed. By contrast, it is the context of the TATA element, rather than its nucleotide sequence that appears to be critical for promoter expression in human cells. Our data suggest the existence of one or more additional factors in human cells that permit the utilization of nonconsensus TATA elements. S.pombe appears to lack these factors.
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U2 - 10.1093/nar/21.11.2737
DO - 10.1093/nar/21.11.2737
M3 - Article
C2 - 8332470
AN - SCOPUS:0027201814
SN - 0305-1048
VL - 21
SP - 2737
EP - 2746
JO - Nucleic acids research
JF - Nucleic acids research
IS - 11
ER -