Nucleic Acids Research, 1989, Vol. 17, No. 22 9205-9218
© 1989
MOLECULAR BIOLOGY |
Characterization of the trancriptional potency of sub-elements of the UAS of the yeast PGK gene in a PGK mini-promoter
Department of Biochemistry, Oxford University South Parks Road, Oxford OX1 3QU 1Department of Molecular Biology British Biotechnology, Watlington Road, Cowley, Oxford OX4 5LY, UK
*To whom correspondence should be addressed
Received July 27, 1989. Revised October 3, 1989. Accepted October 3, 1989.
The upstream activator (UAS) of the yeast PGK gene comprises three different sequence elements. These are 1) a region of strong protein binding called the YFP, 2) three repeats of the motif CTTCC and 3) an essential activator core (AC) sequence that binds the protein RAP1. To assess the function of each of these elements in transcriptional activation we have inserted them individually and in various combinations into a PGK mini-promoter. This comprises only the transcription intiation elements from the PGK promoter and is inactive in the absence of activator sequences. None of the individual sequence elements was capable of activating the mini-promoter. However either the YFP or the CTTCC boxes in conjunction with the AC box resulted in efficient expression. Transcription levels were not however as high as when all three elements were inserted. These data suggest that the efficiency of PGK transcription depends upon the interactions between three different sequences. Furthermore while RAP1 per se is not a transcriptional activator it can associate promiscuously with other factors to create a functional transcription complex.
+Present address: Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DU, UK
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