scholarly journals Fluorescence Spectroscopic Analysis of ppGpp Binding to cAMP Receptor Protein and Histone-Like Nucleoid Structuring Protein

2021 ◽  
Vol 22 (15) ◽  
pp. 7871
Author(s):  
Taner Duysak ◽  
Thanh Tuyen Tran ◽  
Aqeel Rana Afzal ◽  
Che-Hun Jung
Keyword(s):  
Dissociation Constant ◽  
Fluorescence Intensity ◽  
Spectroscopic Analysis ◽  
Dna Packaging ◽  
Intrinsic Fluorescence ◽  
Gene Repression ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Camp Receptor ◽  
Similar Affinity

The cyclic AMP receptor protein (CRP) is one of the best-known transcription factors, regulating about 400 genes. The histone-like nucleoid structuring protein (H-NS) is one of the nucleoid-forming proteins and is responsible for DNA packaging and gene repression in prokaryotes. In this study, the binding of ppGpp to CRP and H-NS was determined by fluorescence spectroscopy. CRP from Escherichia coli exhibited intrinsic fluorescence at 341 nm when excited at 280 nm. The fluorescence intensity decreased in the presence of ppGpp. The dissociation constant of 35 ± 3 µM suggests that ppGpp binds to CRP with a similar affinity to cAMP. H-NS also shows intrinsic fluorescence at 329 nm. The fluorescence intensity was decreased by various ligands and the calculated dissociation constant for ppGpp was 80 ± 11 µM, which suggests that the binding site was occupied fully by ppGpp under starvation conditions. This study suggests the modulatory effects of ppGpp in gene expression regulated by CRP and H-NS. The method described here may be applicable to many other proteins.

scholarly journals The cAMP receptor protein of Trypanosoma cruzi.

1983 ◽  
Vol 258 (11) ◽  
pp. 6979-6983 ◽  
Author(s):  
R Rangel-Aldao ◽  
G Tovar ◽  
M Ledezma de Ruiz
Keyword(s):  
Trypanosoma Cruzi ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Camp Receptor

scholarly journals Interactions between the Escherichia coli cAMP receptor protein and the C-terminal domain of the α subunit of RNA polymerase at Class I promoters

1999 ◽  
Vol 337 (3) ◽  
pp. 415-423 ◽  
Author(s):  
Emma C. LAW ◽  
Nigel J. SAVERY ◽  
Stephen J. W. BUSBY
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Transcription Initiation ◽  
Class I ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Α Subunit ◽  
Terminal Domain ◽  
Up Elements ◽  
Camp Receptor

The Escherichia coli cAMP receptor protein (CRP) is a factor that activates transcription at over 100 target promoters. At Class I CRP-dependent promoters, CRP binds immediately upstream of RNA polymerase and activates transcription by making direct contacts with the C-terminal domain of the RNA polymerase α subunit (αCTD). Since αCTD is also known to interact with DNA sequence elements (known as UP elements), we have constructed a series of semi-synthetic Class I CRP-dependent promoters, carrying both a consensus DNA-binding site for CRP and a UP element at different positions. We previously showed that, at these promoters, the CRP–αCTD interaction and the CRP–UP element interaction contribute independently and additively to transcription initiation. In this study, we show that the two halves of the UP element can function independently, and that, in the presence of the UP element, the best location for the DNA site for CRP is position -69.5. This suggests that, at Class I CRP-dependent promoters where the DNA site for CRP is located at position -61.5, the two αCTDs of RNA polymerase are not optimally positioned. Two experiments to test this hypothesis are presented.

scholarly journals Identification and Characterization of a Novel cAMP Receptor Protein in the CyanobacteriumSynechocystissp. PCC 6803

2000 ◽  
Vol 275 (9) ◽  
pp. 6241-6245 ◽  
Author(s):  
Hidehisa Yoshimura ◽  
Toru Hisabori ◽  
Shuichi Yanagisawa ◽  
Masayuki Ohmori
Keyword(s):  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Camp Receptor ◽  
Identification And Characterization ◽  
Pcc 6803

scholarly journals Regulation of the expression of whiB1 in Mycobacterium tuberculosis: role of cAMP receptor protein

Microbiology ◽  
2006 ◽  
Vol 152 (9) ◽  
pp. 2749-2756 ◽  
Author(s):  
Nisheeth Agarwal ◽  
Tirumalai R. Raghunand ◽  
William R. Bishai
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Binding Site ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Mobility Shift ◽  
Camp Analogue ◽  
Camp Receptor ◽  
Fusion Analysis ◽  
Electrophoretic Mobility Shift Assays

The wbl (whiB-like) genes encode putative transcription factors unique to actinomycetes. This study characterized the promoter element of one of the seven wbl genes of Mycobacterium tuberculosis, whiB1 (Rv3219c). The results reveal that whiB1 is transcribed by a class I-type cAMP receptor protein (CRP)-dependent promoter, harbouring a CRP-binding site positioned at −58.5 with respect to its transcription start point. In vivo promoter activity analysis and electrophoretic mobility shift assays suggest that the expression of whiB1 is indeed regulated by cAMP-dependent binding of CRPM (encoded by the M. tuberculosis gene Rv3676) to the whiB1 5′ untranslated region (5′UTR). β-Galactosidase gene fusion analysis revealed induction of the whiB1 promoter in M. tuberculosis on addition of exogenous dibutyric cAMP (a diffusible cAMP analogue) only when an intact CRP-binding site was present. These results indicate that M. tuberculosis whiB1 transcription is regulated in part by cAMP levels via direct binding of cAMP-activated CRPM to a consensus CRP-binding site in the whiB1 5′UTR.

scholarly journals Thermodynamics of Cyclic Nucleotide Binding to the cAMP Receptor Protein and Its T127L Mutant

1995 ◽  
Vol 270 (37) ◽  
pp. 21679-21683 ◽  
Author(s):  
Inna Gorshkova ◽  
Julie L. Moore ◽  
Keith H. McKenney ◽  
Frederick P. Schwarz
Keyword(s):  
Cyclic Nucleotide ◽  
Nucleotide Binding ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Cyclic Nucleotide Binding ◽  
Camp Receptor
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