TY - JOUR
T1 - Lack of a requirement for strict rotational alignment among transcription factor binding sites in yeast
AU - Inokuchi, Kaoru
AU - Nakayama, Akiko
N1 - Funding Information:
We are most grateful to F. Hishinuma for his encouragment throughout this work. We also thank L. Guarente for plasmid,F. Ozawa for synthesis of oligonucleotides, T. E. Kennedy for critical reading of the manuscript and H. Sakazume for her excellent secretarial assistance. This work was performed as a part of the Research and Development of Basic Technology for Future Industries supported by NEDO of Japan.
PY - 1991/6/11
Y1 - 1991/6/11
N2 - The MFα1 gene of Saccharomyces cerevisiae is an α specific gene whose expression is regulated by two upstream activation sites (UASMFα1s), which are the binding sites for activator proteins, MATα1 and MCM1. UASMFα1 elements act in a synergistic fashion; lack of either element causes reduced expression levels that are 6- to 45-fold less than that of intact promoter in α cells. We investigated the effect of manipulating rotational alignments among transcription factor binding sites of MFα1 on the expression of the gene. The expression level of MFα1 decreased with the spacing between the two UASMFα1s and no synergism was observed when the distance of the two elements was longer than 90 base pairs. No strict rotational alignment was required for their synergistic action. We also show that the spacing between UASMFα1 elements and TFIID binding site, the TATA box, had little, if any, effect on MFα1 expression. We chose a sufficient number of positions for UASMFα1s to ensure that, in several of these positions, MATα1 and MCM1 were on the opposite side of the DNA helix with respect to the TATA box.
AB - The MFα1 gene of Saccharomyces cerevisiae is an α specific gene whose expression is regulated by two upstream activation sites (UASMFα1s), which are the binding sites for activator proteins, MATα1 and MCM1. UASMFα1 elements act in a synergistic fashion; lack of either element causes reduced expression levels that are 6- to 45-fold less than that of intact promoter in α cells. We investigated the effect of manipulating rotational alignments among transcription factor binding sites of MFα1 on the expression of the gene. The expression level of MFα1 decreased with the spacing between the two UASMFα1s and no synergism was observed when the distance of the two elements was longer than 90 base pairs. No strict rotational alignment was required for their synergistic action. We also show that the spacing between UASMFα1 elements and TFIID binding site, the TATA box, had little, if any, effect on MFα1 expression. We chose a sufficient number of positions for UASMFα1s to ensure that, in several of these positions, MATα1 and MCM1 were on the opposite side of the DNA helix with respect to the TATA box.
UR - http://www.scopus.com/inward/record.url?scp=0025905646&partnerID=8YFLogxK
U2 - 10.1093/nar/19.11.3099
DO - 10.1093/nar/19.11.3099
M3 - 学術論文
C2 - 1905400
AN - SCOPUS:0025905646
SN - 0305-1048
VL - 19
SP - 3099
EP - 3103
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 11
ER -