scholarly journals Repression by Cyclic AMP Receptor Protein at a Distance

mBio ◽  
2012 ◽  
Vol 3 (5) ◽  
Author(s):  
David J. Lee ◽  
Stephen J. W. Busby
Keyword(s):  
Escherichia Coli ◽  
Transcription Factors ◽  
Cyclic Amp ◽  
Promoter Activity ◽  
Bacterial Genome ◽  
Receptor Protein ◽  
Transcription Start ◽  
Content Type ◽  
Cyclic Amp Receptor Protein ◽  
Α Subunits

ABSTRACT In a previous study of promoters dependent on the Escherichia coli cyclic AMP receptor protein (CRP), carrying tandem DNA sites for CRP, we found that the upstream-bound CRP could either enhance or repress transcription, depending on its location. Here, we have analyzed the interactions between CRP and the C-terminal domains of the RNA polymerase α subunits at some of these promoters. We report that the upstream-bound CRP interacts with these domains irrespective of whether it up- or downregulates promoter activity. Hence, disruption of this interaction can lead to either down- or upregulation, depending on its location. IMPORTANCE Many bacterial promoters carry multiple DNA sites for transcription factors. While most factors that downregulate promoter activity bind to targets that overlap or are downstream of the transcription start and −10 element, very few cases of repression from upstream locations have been reported. Since more Escherichia coli promoters are regulated by cyclic AMP receptor protein (CRP) than by any other transcription factor, and since multiple DNA sites for CRP are commonplace at promoters, our results suggest that promoter downregulation by transcription factors may be more prevalent than hitherto thought, and this will have implications for the annotation of promoters from new bacterial genome sequences.

scholarly journals Cyclic AMP Receptor Protein RegulatescspE, an Early Cold-Inducible Gene, in Escherichia coli

2011 ◽  
Vol 193 (22) ◽  
pp. 6142-6151 ◽  
Author(s):  
Sheetal Uppal ◽  
Svetlana R. Maurya ◽  
Ramesh S. Hire ◽  
Narendra Jawali
Keyword(s):  
Escherichia Coli ◽  
Low Temperature ◽  
Cyclic Amp ◽  
Target Site ◽  
Class I ◽  
Receptor Protein ◽  
Upstream Region ◽  
Core Motif ◽  
Content Type ◽  
Cyclic Amp Receptor Protein

cspE, a member of thecspAfamily of cold shock proteins inEscherichia coli, is an early cold-inducible protein. The nucleic acid melting ability and transcription antiterminator activity of CspE have been reported to be critical for growth at low temperature. Here, we show that the cyclic AMP receptor protein (CRP), a global regulator involved in sugar metabolism, upregulatescspEinE. coli. Sequence analysis of thecspEupstream region revealed a putative CRP target site centered at −61.5 relative to the transcription start. The binding of CRP to this target site was demonstrated using electrophoretic mobility shift assays. The presence of this site was shown to be essential for PcspEactivation by CRP. Mutational analysis of the binding site indicated that the presence of an intact second core motif is more important than the first core motif for CRP-PcspEinteraction. Based on the promoter architecture, we classified PcspEas a class I CRP-dependent promoter. This was further substantiated by our data demonstrating the involvement of the AR1 domain of CRP in PcspEtranscription. Furthermore, the substitutions in the key residues of the RNA polymerase α-subunit C-terminal domain (α-CTD), which are important for class I CRP-dependent transcription, showed the involvement of 265 and 287 determinants in PcspEtranscription. In addition, the deletion ofcrpled to a growth defect at low temperature, suggesting that CRP plays an important role in cold adaptation.

scholarly journals Binding of the cyclic AMP receptor protein of Escherichia coli and DNA bending at the P4 promoter of pBR322

1992 ◽  
Vol 285 (1) ◽  
pp. 91-97 ◽  
Author(s):  
I Brierley ◽  
J G Hoggett
Keyword(s):  
Escherichia Coli ◽  
Cyclic Amp ◽  
Gel Electrophoresis ◽  
Dna Bending ◽  
Receptor Protein ◽  
Simple Relationship ◽  
Start Site ◽  
Transcription Start ◽  
Bending Angle ◽  
Cyclic Amp Receptor Protein

The binding of the Escherichia coli cyclic AMP receptor protein (CRP) to its specific site on the P4 promoter of pBR322 has been studied by gel electrophoresis. Binding to the P4 site was about 40-50-fold weaker than to the principal CRP site on the lactose promoter at both low (0.01 M) and high (0.1 M) ionic strengths. CRP-induced bending at the P4 site was investigated from the mobilities of CRP bound to circularly permuted P4 fragments. The estimated bending angle, based on comparison with Zinkel & Crothers [(1990) Biopolymers 29, 29-38] A-tract bending standards, was found to be approximately 96 degrees, similar to that found for binding to the lac site. These observations suggest that there is not a simple relationship between strength of CRP binding and the extent of induced bending for different CRP sites. The apparent centre of bending in P4 is displaced about 6-8 bp away from the conserved TGTGA sequence and the P4 transcription start site.

scholarly journals Evidence that Expression of the Vibrio vulnificus Hemolysin Gene Is Dependent on Cyclic AMP and Cyclic AMP Receptor Protein

1999 ◽  
Vol 181 (24) ◽  
pp. 7639-7642 ◽  
Author(s):  
Young Bae Bang ◽  
Shee Eun Lee ◽  
Joon Haeng Rhee ◽  
Sang Ho Choi
Keyword(s):  
Escherichia Coli ◽  
Cyclic Amp ◽  
Promoter Activity ◽  
Vibrio Vulnificus ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Transcriptional Fusion ◽  
Cyclic Amp Receptor Protein ◽  
Hemolysin Gene ◽  
Camp Receptor

ABSTRACT Glucose repressed hemolysin production in Vibrio vulnificus. Promoter activity of the hemolysin gene,vvh, assessed with a vvh-luxCDABEtranscriptional fusion, required cyclic AMP (cAMP) and cAMP receptor protein (CRP) in Escherichia coli. Hemolysin production inV. vulnificus increased after the addition of cAMP and was undetectable in a putative crp mutant, suggesting thatvvh is also regulated by cAMP-CRP in V. vulnificus.

Mannose utilization in Escherichia coli requires cyclic AMP but not an exogenous inducer

1980 ◽  
Vol 26 (12) ◽  
pp. 1508-1511 ◽  
Author(s):  
Ann D. E. Fraser ◽  
Hiroshi Yamazaki
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Cyclic Amp ◽  
Adenylyl Cyclase ◽  
Sole Carbon Source ◽  
Receptor Protein ◽  
Deficient Mutant ◽  
Phosphotransferase System ◽  
Cyclic Amp Receptor Protein ◽  
Cyclic Monophosphate

It has not been clarified whether the utilization of mannose by Escherichia coli requires adenosine 3′,5′-cyclic monophosphate (cyclic AMP). Using an adenylyl cyclase deficient mutant (CA8306B) and a cyclic AMP receptor protein (CRP) deficient mutant (5333B) we have shown that the utilization of mannose is dependent on the cyclic AMP–CRP complex. 2-Deoxyglucose (DG) is a nonmetabolizable glucose analog specific for the phosphotransferase system (PTS) which transports mannose (termed here PTSM). Growth of CA8306B on glycerol is unaffected by addition of the analog, whereas growth of the strain on glycerol plus cyclic AMP ceases im mediately upon addition of DG. These results suggest that the formation of PTSM is dependent on cyclic AMP. In addition, CA8306B grown on glycerol plus cyclic AMP can immediately utilize mannose when transferred to a medium containing mannose as a sole carbon source, whereas the same strain grown on glycerol without cyclic AMP cannot utilize mannose when so transferred. These results suggest that the formation of PTSM does not require an exogenous inducer.

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