Functional analysis of different sequence elements in the Escherichia coli galactose operon P 2 promoter
- 1 March 1988
- journal article
- research article
- Published by Wiley in Molecular Microbiology
- Vol. 2 (2) , 165-172
- https://doi.org/10.1111/j.1365-2958.1988.tb00018.x
Abstract
Starting with a DNA fragment containing the galactose operon P2 promoter, we made a series of deletions that progressively replaced DNA sequences upstream of the transcription startpoint and determined their effects on P2 activity. The results show that specific sequences upstream of -32 are not important. Removal of the sequence 5''-CACA-3'' from -32 to -28 reduces P2 activity by 50%: longer deletions to -16 further reduce activity but do not remove the information specifying the transcription startpoint. DNA sequences between -32 and -16 at gal P2 assist the isomerization of RNA polymerase from closed to open complexes rather than contributing to the initial binding of RNA polymerase. The activity of gal P2 in the absence of -35 region sequences is dependent on the sequence TG just upstream of the -10 hexamer, TATACT: a mutation at -14 changing the TG sequence to TT totally inactivates P2. However, P2 activity can be restored if the consensus -35 sequence TTGACA is cloned 17 bp upstream of the -10 hexamer. Thus, for transcription initiation, the -10 hexamer, TATACT, must ''cooperate'' with upstream sequences that may be located either around -35 or -14.This publication has 26 references indexed in Scilit:
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