scholarly journals Sycrp2 Is Essential for Twitching Motility in the CyanobacteriumSynechocystissp. Strain PCC 6803

2018 ◽  
Vol 200 (21) ◽  
Author(s):  
Wei-Yu Song ◽  
Sha-Sha Zang ◽  
Zheng-Ke Li ◽  
Guo-Zheng Dai ◽  
Ke Liu ◽  
...  
Keyword(s):  
Cell Motility ◽  
Receptor Protein ◽  
Upstream Region ◽  
Twitching Motility ◽  
Camp Receptor Protein ◽  
Mobility Shift ◽  
Content Type ◽  
Receptor Proteins ◽  
Camp Receptor ◽  
Pcc 6803

ABSTRACTTwo cAMP receptor proteins (CRPs), Sycrp1 (encoded bysll1371) and Sycrp2 (encoded bysll1924), exist in the cyanobacteriumSynechocystissp. strain PCC 6803. Previous studies have demonstrated that Sycrp1 has binding affinity for cAMP and is involved in motility by regulating the formation of pili. However, the function of Sycrp2 remains unknown. Here, we report thatsycrp2disruption results in the loss of motility ofSynechocystissp. PCC 6803, and that the phenotype can be recovered by reintroducing thesycrp2gene into the genome ofsycrp2-disrupted mutants. Electron microscopy showed that thesycrp2-disrupted mutant lost the pilus apparatus on the cell surface, resulting in a lack of cell motility. Furthermore, the transcript level of thepilA9-pilA11operon (essential for cell motility and regulated by the cAMP receptor protein Sycrp1) was markedly decreased insycrp2-disrupted mutants compared with the wild-type strain. Moreover, yeast two-hybrid analysis and a pulldown assay demonstrated that Sycrp2 interacted with Sycrp1 to form a heterodimer and that Sycrp1 and Sycrp2 interacted with themselves to form homodimers. Gel mobility shift assays revealed that Sycrp1 specifically binds to the upstream region ofpilA9. Together, these findings indicate that inSynechocystissp. PCC 6803, Sycrp2 regulates the formation of pili and cell motility by interacting with Sycrp1.IMPORTANCEcAMP receptor proteins (CRPs) are widely distributed in cyanobacteria and play important roles in regulating gene expression. Although many cyanobacterial species have two cAMP receptor-like proteins, the functional links between them are unknown. Here, we found that Sycrp2 in the cyanobacteriumSynechocystissp. strain PCC 6803 is essential for twitching motility and that it interacts with Sycrp1, a known cAMP receptor protein involved with twitching motility. Our findings indicate that the two cAMP receptor-like proteins in cyanobacteria do not have functional redundancy but rather work together.

scholarly journals A cAMP Receptor Protein, SYCRP1, is Responsible for the Cell Motility of Synechocystis sp. PCC 6803

2002 ◽  
Vol 43 (4) ◽  
pp. 460-463 ◽  
Author(s):  
Hidehisa Yoshimura ◽  
Shizue Yoshihara ◽  
Shinobu Okamoto ◽  
Masahiko Ikeuchi ◽  
Masayuki Ohmori
Keyword(s):  
Cell Motility ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Camp Receptor ◽  
Synechocystis Sp ◽  
Pcc 6803

scholarly journals Cyclic AMP (cAMP) Receptor Protein-cAMP Complex Regulates Heparosan Production in Escherichia coli Strain Nissle 1917

2015 ◽  
Vol 81 (22) ◽  
pp. 7687-7696 ◽  
Author(s):  
Huihui Yan ◽  
Feifei Bao ◽  
Liping Zhao ◽  
Yanying Yu ◽  
Jiaqin Tang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cyclic Amp ◽  
Carbon Sources ◽  
Coli Strain ◽  
Site Directed Mutagenesis ◽  
Receptor Protein ◽  
Upstream Region ◽  
Camp Receptor Protein ◽  
Content Type ◽  
Camp Receptor

ABSTRACTHeparosan serves as the starting carbon backbone for the chemoenzymatic synthesis of heparin, a widely used clinical anticoagulant drug. The availability of heparosan is a significant concern for the cost-effective synthesis of bioengineered heparin. The carbon source is known as the pivotal factor affecting heparosan production. However, the mechanism by which carbon sources control the biosynthesis of heparosan is unclear. In this study, we found that the biosynthesis of heparosan was influenced by different carbon sources. Glucose inhibits the biosynthesis of heparosan, while the addition of either fructose or mannose increases the yield of heparosan. Further study demonstrated that the cyclic AMP (cAMP)-cAMP receptor protein (CRP) complex binds to the upstream region of the region 3 promoter and stimulates the transcription of the gene cluster for heparosan biosynthesis. Site-directed mutagenesis of the CRP binding site abolished its capability of binding CRP and eliminated the stimulative effect on transcription.1H nuclear magnetic resonance (NMR) analysis was further performed to determine theEscherichia colistrain Nissle 1917 (EcN) heparosan structure and quantify extracellular heparosan production. Our results add to the understanding of the regulation of heparosan biosynthesis and may contribute to the study of other exopolysaccharide-producing strains.

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 Repression of VvpM Protease Expression by Quorum Sensing and the cAMP-cAMP Receptor Protein Complex inVibrio vulnificus

2018 ◽  
Vol 200 (7) ◽  
Author(s):  
Jeong-A Kim ◽  
Mi-Ae Lee ◽  
You-Chul Jung ◽  
Bo-Ram Jang ◽  
Kyu-Ho Lee
Keyword(s):  
Transcription Factors ◽  
Stationary Phase ◽  
Cell Density ◽  
Transcription Initiation ◽  
Vibrio Vulnificus ◽  
The Other ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Content Type ◽  
Camp Receptor

ABSTRACTSepticemia-causingVibrio vulnificusproduces at least three exoproteases, VvpE, VvpS, and VvpM, all of which participate in interactions with human cells. Expression of VvpE and VvpS is induced in the stationary phase by multiple transcription factors, including sigma factor S, SmcR, and the cAMP-cAMP receptor protein (cAMP-CRP) complex. Distinct roles of VvpM, such as induction of apoptosis, lead us to hypothesize VvpM expression is different from that of the other exoproteases. Its transcription, which was found to be independent of sigma S, is induced at the early exponential phase and then becomes negligible upon entry into the stationary phase. SmcR and CRP were studied regarding the control ofvvpMexpression. Transcription ofvvpMwas repressed by SmcR and cAMP-CRP complex individually, which specifically bound to the regions −2 to +20 and +6 to +27, respectively, relative to thevvpMtranscription initiation site. Derepression ofvvpMgene expression was 10- to 40-fold greater in ansmcR crpdouble mutant than in single-gene mutants. Therefore, these results show that the expression ofV. vulnificusexoproteases is differentially regulated, and in this way, distinct proteases can engage in specific interactions with a host.IMPORTANCEAn opportunistic human pathogen,Vibrio vulnificusproduces multiple extracellular proteases that are involved in diverse interactions with a host. The total exoproteolytic activity is detected mainly in the supernatants of the high-cell-density cultures. However, some proteolytic activity derived from a metalloprotease, VvpM, was present in the supernatants of the low-cell-density cultures sampled at the early growth period. In this study, we present the regulatory mechanism for VvpM expression via repression by at least two transcription factors. This type of transcriptional regulation is the exact opposite of those for expression of the otherV. vulnificusexoproteases. Differential regulation of each exoprotease's production then facilitates the pathogen's participation in the distinct interactions with a host.

scholarly journals Systematic single base-pair substitution analysis of DNA binding by the cAMP receptor protein in cyanobacteriumSynechocystissp. PCC 6803

FEBS Letters ◽  
2004 ◽  
Vol 563 (1-3) ◽  
pp. 55-58 ◽  
Author(s):  
Katsumi Omagari ◽  
Hidehisa Yoshimura ◽  
Mitsunori Takano ◽  
Dongyun Hao ◽  
Masayuki Ohmori ◽  
...  
Keyword(s):  
Dna Binding ◽  
Base Pair ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Single Base ◽  
Single Base Pair Substitution ◽  
Single Base Pair ◽  
Base Pair Substitution ◽  
Camp Receptor ◽  
Pcc 6803

scholarly journals Positive Effect of Carbon Sources on Natural Transformation in Escherichia coli: Role of Low-Level Cyclic AMP (cAMP)-cAMP Receptor Protein in the Derepression ofrpoS

2015 ◽  
Vol 197 (20) ◽  
pp. 3317-3328 ◽  
Author(s):  
Mengyue Guo ◽  
Huanyu Wang ◽  
Nengbin Xie ◽  
Zhixiong Xie
Keyword(s):  
Escherichia Coli ◽  
Natural Transformation ◽  
Carbon Sources ◽  
Transformation Frequency ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Content Type ◽  
E Coli ◽  
Camp Receptor

ABSTRACTNatural plasmid transformation ofEscherichia coliis a complex process that occurs strictly on agar plates and requires the global stress response factor σS. Here, we showed that additional carbon sources could significantly enhance the transformability ofE. coli. Inactivation of phosphotransferase system genes (ptsH,ptsG, andcrr) caused an increase in the transformation frequency, and the addition of cyclic AMP (cAMP) neutralized the promotional effect of carbon sources. This implies a negative role of cAMP in natural transformation. Further study showed thatcrpandcyaAmutations conferred a higher transformation frequency, suggesting that the cAMP-cAMP receptor protein (CRP) complex has an inhibitory effect on transformation. Moreover, we observed thatrpoSis negatively regulated by cAMP-CRP in early log phase and that bothcrpandcyaAmutants show no transformation superiority whenrpoSis knocked out. Therefore, it can be concluded that both thecrpandcyaAmutations derepressrpoSexpression in early log phase, whereby they aid in the promotion of natural transformation ability. We also showed that the accumulation of RpoS during early log phase can account for the enhanced transformation aroused by additional carbon sources. Our results thus demonstrated that the presence of additional carbon sources promotes competence development and natural transformation by reducing cAMP-CRP and, thus, derepressingrpoSexpression during log phase. This finding could contribute to a better understanding of the relationship between nutrition state and competence, as well as the mechanism of natural plasmid transformation inE. coli.IMPORTANCEEscherichia coli, which is not usually considered to be naturally transformable, was found to spontaneously take up plasmid DNA on agar plates. Researching the mechanism of natural transformation is important for understanding the role of transformation in evolution, as well as in the transfer of pathogenicity and antibiotic resistance genes. In this work, we found that carbon sources significantly improve transformation by decreasing cAMP. Then, the low level of cAMP-CRP derepresses the general stress response regulator RpoS via a biphasic regulatory pattern, thereby contributing to transformation. Thus, we demonstrate the mechanism by which carbon sources affect natural transformation, which is important for revealing information about the interplay between nutrition state and competence development inE. coli.

scholarly journals cAMP Receptor Protein ControlsVibrio choleraeGene Expression in Response to Host Colonization

mBio ◽  
2018 ◽  
Vol 9 (4) ◽  
Author(s):  
Jainaba Manneh-Roussel ◽  
James R. J. Haycocks ◽  
Andrés Magán ◽  
Nicolas Perez-Soto ◽  
Kerstin Voelz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Vibrio Cholerae ◽  
Expression Patterns ◽  
Lifestyle Changes ◽  
Receptor Protein ◽  
Camp Receptor Protein ◽  
Host Colonization ◽  
Content Type ◽  
Camp Receptor

ABSTRACTThe bacteriumVibrio choleraeis native to aquatic environments and can switch lifestyles to cause disease in humans. Lifestyle switching requires modulation of genetic systems for quorum sensing, intestinal colonization, and toxin production. Much of this regulation occurs at the level of gene expression and is controlled by transcription factors. In this work, we have mapped the binding of cAMP receptor protein (CRP) and RNA polymerase across theV. choleraegenome. We show that CRP is an integral component of the regulatory network that controls lifestyle switching. Focusing on a locus necessary for toxin transport, we demonstrate CRP-dependent regulation of gene expression in response to host colonization. Examination of further CRP-targeted genes reveals that this behavior is commonplace. Hence, CRP is a key regulator of manyV. choleraegenes in response to lifestyle changes.IMPORTANCECholera is an infectious disease that is caused by the bacteriumVibrio cholerae. Best known for causing disease in humans, the bacterium is most commonly found in aquatic ecosystems. Hence, humans acquire cholera following ingestion of food or water contaminated withV. cholerae. Transition between an aquatic environment and a human host triggers a lifestyle switch that involves reprogramming ofV. choleraegene expression patterns. This process is controlled by a network of transcription factors. In this paper, we show that the cAMP receptor protein (CRP) is a key regulator ofV. choleraegene expression in response to lifestyle changes.

scholarly journals Mutations That StimulateflhDCExpression in Escherichia coli K-12

2015 ◽  
Vol 197 (19) ◽  
pp. 3087-3096 ◽  
Author(s):  
Karen A. Fahrner ◽  
Howard C. Berg
Keyword(s):  
Escherichia Coli ◽  
Transcription Factors ◽  
Insertion Sequence ◽  
Receptor Protein ◽  
Upstream Region ◽  
Single Nucleotide ◽  
Content Type ◽  
Sequence Elements ◽  
Camp Receptor ◽  
K 12

ABSTRACTMotility is a beneficial attribute that enables cells to access and explore new environments and to escape detrimental ones. The organelle of motility inEscherichia coliis the flagellum, and its production is initiated by the activating transcription factors FlhD and FlhC. The expression of these factors by theflhDCoperon is highly regulated and influenced by environmental conditions. TheflhDCpromoter is recognized by σ70and is dependent on the transcriptional activator cyclic AMP (cAMP)-cAMP receptor protein complex (cAMP-CRP). A number of K-12 strains exhibit limited motility due to low expression levels offlhDC. We report here a large number of mutations that stimulateflhDCexpression in such strains. They include single nucleotide changes in the −10 element of the promoter, in the promoter spacer, and in the cAMP-CRP binding region. In addition, we show that insertion sequence (IS) elements or a kanamycin gene located hundreds of base pairs upstream of the promoter can effectively enhance transcription, suggesting that the topology of a large upstream region plays a significant role in the regulation offlhDCexpression. None of the mutations eliminated the requirement for cAMP-CRP for activation. However, several mutations allowed expression in the absence of the nucleoid organizing protein, H-NS, which is normally required forflhDCexpression.IMPORTANCETheflhDCoperon ofEscherichia coliencodes transcription factors that initiate flagellar synthesis, an energetically costly process that is highly regulated. Few deregulating mutations have been reported thus far. This paper describes new single nucleotide mutations that stimulateflhDCexpression, including a number that map to the promoter spacer region. In addition, this work shows that insertion sequence elements or a kanamycin gene located far upstream from the promoter or repressor binding sites also stimulate transcription, indicating a role of regional topology in the regulation offlhDCexpression.

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