Transcriptional regulation of the Enterobacter cloacae UW4 1-aminocyclopropane-1-carboxylate (ACC) deaminase gene (acdS)

2001 ◽  
Vol 47 (4) ◽  
pp. 359-367 ◽  
Author(s):  
Jiping Li ◽  
Bernard R Glick
Keyword(s):  
Transcriptional Regulation ◽  
Luciferase Activity ◽  
Growth Promotion ◽  
Regulatory Protein ◽  
Acc Deaminase ◽  
Enterobacter Cloacae ◽  
Upstream Region ◽  
Luciferase Expression ◽  
Luciferase Gene ◽  
The One

Based on DNA sequence analysis and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, the region of DNA immediately upstream of the Enterobacter cloacae UW4 ACC deaminase gene (acdS) contains several features that appear to be involved in its transcriptional regulation. In the present study, the 5' upstream region of acdS was cloned into the promoter-probe vector, pQF70, which carries the promoterless luciferase gene (luxAB), and luciferase expression was monitored. The data obtained from studying the expression of the luciferase gene showed that (i) a leucine responsive regulatory protein (LRP)-like protein encoded within the upstream region is located on the opposite strand from acdS under the control of a promoter stronger than the one responsible for acdS transcription, (ii) luciferase gene expression required both ACC and the LRP-like protein, (iii) luciferase expression was increased three-fold under anaerobic conditions, consistent with the involvement of a fumarate-nitrate reduction (FNR)-like regulatory protein box within the upstream region, and (iv) the addition of leucine to the growth medium decreased luciferase activity in the presence of ACC and increased luciferase activity in the absence of ACC, consistent with leucine acting as a regulator of the expression of the LRP-like protein.Key words: plant growth promotion, ethylene, ACC deaminase, regulation, Enterobacter cloacae.

Facilitated DNA transfer to rat submandibular gland in vivo and GRP-Ca gene regulation

1995 ◽  
Vol 268 (6) ◽  
pp. G1074-G1078 ◽  
Author(s):  
B. C. O'Connell ◽  
K. G. Ten Hagen ◽  
K. W. Lazowski ◽  
L. A. Tabak ◽  
B. J. Baum
Keyword(s):  
Luciferase Activity ◽  
Firefly Luciferase ◽  
Upstream Region ◽  
Luciferase Expression ◽  
Upstream Sequence ◽  
Luciferase Gene ◽  
Base Pairs ◽  
Firefly Luciferase Gene ◽  
Rat Submandibular Gland

The internalization of DNA can be facilitated by adenovirus infection. Using the replication-deficient adenovirus, Ad-dl312, and a plasmid-based firefly luciferase gene as a reporter, we have optimized the uptake and expression of DNA in rat submandibular glands in vivo. Luciferase expression is transient and peaked at approximately 18 h after infection. Luciferase activity increased with plasmid concentration and was greatest at 10(9) to 10(10) plaque-forming units of Ad-dl312 per gland. We next examined the expression in vivo of plasmids containing deletions of the glutamine/glutamic acid-rich protein (GRP-Ca isoform) gene upstream region linked to a chloramphenicol acetyltransferase (CAT) reporter. Constructs with 9.4, 6.3, and 2.7 kb and 17 base pairs of upstream sequence gave relative CAT activities of 100, 30, 7.6, and 38.5, respectively. With the 9.4-kb GRP-Ca construct, CAT was preferentially expressed in acinar cells, which is characteristic of GRP. This gene transfer approach should prove useful in the further study of gene expression in salivary glands and other organs.

scholarly journals Aromatic Amino Acid-Dependent Expression of Indole-3-Pyruvate Decarboxylase Is Regulated by TyrR in Enterobacter cloacae UW5

2008 ◽  
Vol 190 (21) ◽  
pp. 7200-7208 ◽  
Author(s):  
R. Julie Ryu ◽  
Cheryl L. Patten
Keyword(s):  
Amino Acid ◽  
Plant Growth ◽  
Aromatic Amino Acid ◽  
Growth Promotion ◽  
Regulatory Protein ◽  
Pyruvate Decarboxylase ◽  
Enterobacter Cloacae ◽  
Acid Uptake ◽  
Mobility Shift ◽  
Iaa Production

ABSTRACT The plant growth-promoting rhizobacterium Enterobacter cloacae UW5 synthesizes the plant growth hormone indole-3-acetic acid (IAA) via the indole-3-pyruvate pathway utilizing the enzyme indole-3-pyruvate decarboxylase that is encoded by ipdC. In this bacterium, ipdC expression and IAA production occur in stationary phase and are induced by an exogenous source of tryptophan, conditions that are present in the rhizosphere. The aim of this study was to identify the regulatory protein that controls the expression of ipdC. We identified a sequence in the promoter region of ipdC that is highly similar to the recognition sequence for the Escherichia coli regulatory protein TyrR that regulates genes involved in aromatic amino acid transport and metabolism. Using a tyrR insertional mutant, we demonstrate that TyrR is required for IAA production and for induction of ipdC transcription. TyrR directly induces ipdC expression, as was determined by real-time quantitative reverse transcription-PCR, by ipdC promoter-driven reporter gene activity, and by electrophoretic mobility shift assays. Expression increases in response to tryptophan, phenylalanine, and tyrosine. This suggests that, in addition to its function in plant growth promotion, indolepyruvate decarboxylase may be important for aromatic amino acid uptake and/or metabolism.

Identification of DNA sequences that regulate the expression of the Enterobacter cloacae UW4 1-aminocyclopropane-1-carboxylic acid deaminase gene

2000 ◽  
Vol 46 (12) ◽  
pp. 1159-1165 ◽  
Author(s):  
Varvara P Grichko ◽  
Bernard R Glick
Keyword(s):  
Protein Binding ◽  
Binding Site ◽  
Dna Sequences ◽  
Regulatory Protein ◽  
Acc Deaminase ◽  
Enterobacter Cloacae ◽  
Receptor Protein ◽  
Protein Binding Site ◽  
Coding Region ◽  
Bacterial Strains

Analysis of the DNA sequence upstream of the previously isolated Enterobacter cloacae UW4 ACC deaminase gene (Shah et al. 1998) suggests that this segment contains several features that are thought to be involved in the transcriptional regulation of this gene. These features include half of a CRP (cAMP receptor protein) binding site, an FNR (fumarate-nitrate reduction) regulatory protein binding site, an LRP (leucine responsive regulatory protein) binding site, and an LRP-like protein coding region. ACC deaminase activity was measured following growth of either various Escherichia coli strains carrying a plasmid that contained the Enterobacter cloacae UW4 ACC deaminase gene, or of Enterobacter cloacae UW4. Variables that were compared include aerobic versus anaerobic conditions, the presence and absence of ACC in the growth medium, addition of leucine to the medium, and bacterial strains that did or did not contain either lrp or fnr genes. The data reported are consistent with the involvement of most, if not all, of the above mentioned potential regulatory regions in the expression of ACC deaminase.Key words: 1-aminocyclopropane-1-carboxylate, ACC, plant growth-promoting rhizobacteria, ACC deaminase, Enterobacter cloacae, leucine responsive regulatory protein.

scholarly journals Screening, Identification and Growth-Promotion Products of Multifunctional Bacteria in a Chinese Fir Plantation

Forests ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 120
Author(s):  
Guangyu Zhao ◽  
Yihui Wei ◽  
Jiaqi Chen ◽  
Yuhong Dong ◽  
Lingyu Hou ◽  
...  
Keyword(s):  
High Efficiency ◽  
Growth Promotion ◽  
Nitrogenase Activity ◽  
Acc Deaminase ◽  
Inorganic Phosphorus ◽  
Chinese Fir ◽  
Biochemical Tests ◽  
Growth Promoting ◽  
Chinese Fir Plantation ◽  
Physiological And Biochemical

Purpose: This research was aimed to screen and identify multifunctional phosphorus-dissolving bacteria of a Chinese fir (Cunninghamia lanceolata) plantation and study its phosphorus-dissolving characteristics in order to provide strain resources and a theoretical basis for developing the appropriate bacterial fertilizer of a Chinese fir plantation. Methods: First, phosphorus-dissolving bacteria were isolated from the woodland soil of a Chinese fir plantation by Pikovskava inorganic phosphorus medium (PVK). Then, some growth-promoting indicators of primary screening strains were determined, including the capacity of phosphorus-solubilized, nitrogenase activity, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, production of indole-3-acetic acid (IAA), secretion of iron carrier and so on. Finally, the screening multifunctional phosphorus-dissolving bacteria were identified, which were combined with colony characteristics, physiological and biochemical tests and molecular biotechnology. Results: (1) Thirteen phosphorus-dissolving bacteria were isolated and screened in total, and P5 (195.61 mg·L−1) had the strongest capacity of phosphorus-solubilized. Five phosphorus-dissolving bacteria were provided with nitrogenase activity, and the highest activity of nitrogenase was P10 and P5 (71.90 C2H4 nmol·mL−1·h−1 and 71.00 C2H4 nmol·mL−1·h−1, respectively). Four strains were provided with ACC deaminase activity, and the highest activity of ACC deaminase was P5 and P9, (0.74 μmol·mg−1·h−1 and 0.54 μmol·mg−1·h−1, respectively). Most strains could secrete IAA, and three strains of bacteria had a strong secretory ability, which could secrete IAA with a concentration greater than 15 mg·mL−1, and P5 was 18.00, P2 was 17.30, P6 was 15.59 (mg·mL−1). P5 produced carriers of iron better than others, and the ratio of the diameter of the iron production carrier ring to the diameter of the colony was 1.80, respectively, which was significantly higher than other strains. Combining all kinds of factors, P5 multifunctional phosphorus-dissolving bacteria were screened for eventual further study. (2) Strain P5 was identified as Burkholderia ubonensis, based on the colony characteristics, physiological and biochemical tests, 16SrDNA sequence analysis and phylogenetic tree construction. Conclusion: P5 has a variety of high-efficiency growth-promoting capabilities, and the ability to produce IAA, ACC deaminase activity and siderophore performance are significantly higher than other strains, which had great potential in the development of microbial fertilizer.

Transcriptional regulation of Pseudomonas aeruginosa rhlR, encoding a quorum-sensing regulatory protein

Microbiology ◽  
2003 ◽  
Vol 149 (11) ◽  
pp. 3073-3081 ◽  
Author(s):  
Gerardo Medina ◽  
Katy Juárez ◽  
Rafael Díaz ◽  
Gloria Soberón-Chávez
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Transcriptional Regulation ◽  
Quorum Sensing ◽  
Regulatory Protein ◽  
Culture Conditions ◽  
Transcriptional Level ◽  
Coding Region ◽  
Upstream Gene ◽  
Transcription Start Sites ◽  
Response Systems

The Pseudomonas aeruginosa rhlR gene encodes the transcriptional regulator RhlR which has a central role in the quorum-sensing response. Different gene products involved in bacterial pathogenesis are regulated at the transcriptional level by two quorum-sensing response systems, Las and Rhl. The expression of rhlR has been reported to be under the control of the Las system, but its transcriptional regulation has not been studied in detail. Here, the rhlR promoter region has been characterized and shown to present four different transcription start sites, two of which are included in the upstream gene (rhlB) coding region. It was found that rhlR expression is not only dependent on LasR but also on different regulatory proteins such as Vfr and RhlR itself, and also on the alternative sigma factor σ 54. It is reported that rhlR expression is partially LasR-independent under certain culture conditions and is strongly influenced by environmental factors.

scholarly journals Quorum Sensing System Affects the Plant Growth Promotion Traits of Serratia fonticola GS2

2020 ◽  
Vol 11 ◽  
Author(s):  
Byung Kwon Jung ◽  
Jerald Conrad Ibal ◽  
Huy Quang Pham ◽  
Min-Chul Kim ◽  
Gun-Seok Park ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Plant Growth ◽  
Plant Growth Promotion ◽  
Biofilm Formation ◽  
Growth Promotion ◽  
Acc Deaminase ◽  
Deletion Mutants ◽  
Phenotypic Data ◽  
Collective Behaviors ◽  
Pgp Activities

Quorum sensing (QS) enables bacteria to organize gene expression programs, thereby coordinating collective behaviors. It involves the production, release, and population-wide detection of extracellular signaling molecules. The cellular processes regulated by QS in bacteria are diverse and may be used in mutualistic coordination or in response to changing environmental conditions. Here, we focused on the influence of the QS-dependent genes of our model bacterial strain Serratia fonticola GS2 on potential plant growth promoting (PGP) activities including indole-3-acetic acid (IAA) production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, and biofilm formation. Based on genomic and phenotypic experimental data we identified and investigated the function of QS genes in the genome of the model strain. Our gene deletion study confirmed the biological functionality of the QS auto-inducer (gloI) and receptor (gloR) on potential PGP activities of GS2. A transcriptomic approach was also undertaken to understand the role of QS genes in regulation of genes primarily involved in PGP activities (IAA, ACC deaminase activity, and biofilm formation). Both transcriptomic and phenotypic data revealed that the QS-deletion mutants had considerably less PGP activities, as compared to the wild type. In addition, in vivo plant experiments showed that plants treated with GS2 had significantly higher growth rates than plants treated with the QS-deletion mutants. Overall, our results showed how QS-dependent genes regulate the potential PGP activities of GS2. This information may be helpful in understanding the relationship between QS-dependent genes and the PGP activity of bacteria, which aid in the production of practical bio-fertilizers for plant growth promotion.

scholarly journals Topoisomerase activity is linked to altered nucleosome positioning and transcriptional regulation in the fission yeast fbp1 gene

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0242348
Author(s):  
Ryuta Asada ◽  
Satoshi Senmatsu ◽  
Ben Montpetit ◽  
Kouji Hirota
Keyword(s):  
Transcriptional Regulation ◽  
Fission Yeast ◽  
Chromatin Structure ◽  
Promoter Region ◽  
Nucleosome Positioning ◽  
Dna Topology ◽  
Upstream Region ◽  
Inactive Form ◽  
Topological State ◽  
Transcriptional Output

Chromatin structure, including nucleosome positioning, has a fundamental role in transcriptional regulation through influencing protein-DNA interactions. DNA topology is known to influence chromatin structure, and in doing so, can also alter transcription. However, detailed mechanism(s) linking transcriptional regulation events to chromatin structure that is regulated by changes in DNA topology remain to be well defined. Here we demonstrate that nucleosome positioning and transcriptional output from the fission yeast fbp1 and prp3 genes are altered by excess topoisomerase activity. Given that lncRNAs (long noncoding RNAs) are transcribed from the fbp1 upstream region and are important for fbp1 gene expression, we hypothesized that local changes in DNA topological state caused by topoisomerase activity could alter lncRNA and fbp1 transcription. In support of this, we found that topoisomerase overexpression caused destabilization of positioned nucleosomes within the fbp1 promoter region, which was accompanied by aberrant fbp1 transcription. Similarly, the direct recruitment of topoisomerase, but not a catalytically inactive form, to the promoter region of fbp1 caused local changes in nucleosome positioning that was also accompanied by altered fbp1 transcription. These data indicate that changes in DNA topological state induced by topoisomerase activity could lead to altered fbp1 transcription through modulating nucleosome positioning.

scholarly journals Exploitation of endophytic Pseudomonas sp. for plant growth promotion and colonization in rice

2017 ◽  
Vol 9 (3) ◽  
pp. 1310-1316
Author(s):  
Gurjot Kaur ◽  
Poonam Sharma ◽  
Deepika Chhabra ◽  
Kailash Chand ◽  
Gurjit Singh Mangat
Keyword(s):  
Plant Growth ◽  
Plant Growth Promotion ◽  
Growth Promotion ◽  
Acc Deaminase ◽  
Pseudomonas Sp ◽  
Bacterial Isolates ◽  
Pgp Traits ◽  
P Solubilization ◽  
Fluorescent Pigment

The present investigation was carried out to exploit bacterial endophytes associated with root and leaf tissue of rice plant for plant growth promotion (PGP) and colonization study in vitro. Total 10 endophytic bacterial isolates (Pseudomonas sp.) were evaluate for PGP traits like P solubilization, production of Indole acetic acid (IAA), siderophore, ACC deaminase, protease, cellulase, fluorescent pigment, urease and denitrification activity. Out of 10 endophytic bacteria 30 %, 60 %, 20 %, 70 %, 10 % and 10 % were positive for siderophore, protease, cellulase, fluorescent pigment, urease and denitrification respectively. Maximum IAA production was recorded with isolate LRBLE7 (18.8 μgml-1) followed by LRBRE4 (16.0 μgml-1) and maximum P-solubilization was recorded with isolate LRBRE4 (5.8 mg 100 ml-1) followed by LRBLE7 (4.4 mg 100 ml-1). ACC deaminase production was recorded with isolate LRBLE6 (O.D=0.352 nm) followed by LRBRE5 (O.D=0.324nm). Three potential isolates (LRBRE4, LRBRE6 and LRBLE7) were selected on the basis of multiple PGP traits and were subjected to colonization study of rice seedling in vitro. Potential bacterial isolates can be exploited for improving growth and productivity in rice under sustainable management system.

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