Characterization of promoter recognition complexes formed by CRP and CytR for repression and by CRP and RNA polymerase for activation of transcription on the Escherichia coli deoP2 promoter.
The structure of the cAMP-CRP-CytR repression complex and the cAMP CRP-RNA polymerase initiation complex at the deoP2 promoter of E. coli have been probed by DNase I and uranyl footprinting. In the CRP2-CytR complex all protein DNA-phosphate contacts at CRP-1 and CRP-2 are retained, and in addition two new minor groove contacts, ascribed to phosphate-CytR interactions, are observed at -60 between the CRP sites. The contacts are compatible with a model in which the promoter DNA is wrapped around a complex of two CRPs and one CytR. In the RNA polymerase-CRP complex, the CRP-1 phosphate contacts are almost identical to those seen in the repression complex and strong RNA polymerase contacts are seen in the -10 and in the +10 regions. Most noteworthy are minor groove contacts in the -60 region ascribed to RNA polymerase contacts upstream from the CRP. Furthermore, binding of CRP to the CRP-2 target does not seem to interfere with RNA polymerase binding. Thus, a model is suggested in which the DNA is wrapped around a complex of RNA polymerase and one CRP. Finally, the results show that CytR and RNA polymerase are rivals that compete for binding with CRP at deoP2 and that CytR functions as an antiactivator.
Møllegaard
N E
NE
Department of Biochemistry B, Panum Institute, University of Copenhagen, Denmark.
Rasmussen
P B
PB
Valentin-Hansen
P
P
Nielsen
P E
PE
D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein; D004269:DNA, Bacterial / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D003850:Deoxyribonuclease I; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011953:Receptors, Cyclic AMP / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D013696:Temperature; D015533:Transcriptional Activation
D001426:Bacterial Proteins; D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; C048425:CytR protein, E coli; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D011953:Receptors, Cyclic AMP; D012097:Repressor Proteins; D000242:Cyclic AMP; D012321:DNA-Directed RNA Polymerases; D003850:Deoxyribonuclease I