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5 results on '"GAC-RSM"'

1. Expression, regulation and physiological roles of the five Rsm proteins in Pseudomonas syringae pv. tomato DC3000.

Author

Vásquez, Adriana, Ferreiro, María-Dolores, Martínez-Rodríguez, Laura, and Gallegos, María-Trinidad

Subjects
*GENE expression, *PSEUDOMONAS syringae, *RNA-binding proteins, *GRAM-positive bacteria, *SECONDARY metabolism
Abstract

Proteins belonging to the RsmA (regulator of secondary metabolism)/CsrA (carbon storage regulator) family are small RNA-binding proteins that play crucial roles post-transcriptionally regulating gene expression in many Gram-negative and some Gram-positive bacteria. Although most of the bacteria studied have a single RsmA/CsrA gene, Pseudomonas syringae pv. tomato (Pto) DC3000 encodes five Rsm proteins: RsmA/CsrA2, RsmC/CsrA1, RsmD/CsrA4, RsmE/CsrA3, and RsmH/CsrA5. This work aims to provide a comprehensive analysis of the expression of these five rsm protein-encoding genes, elucidate the regulatory mechanisms governing their expression, as well as the physiological relevance of each variant. To achieve this, we examined the expression of rsmA, rsmE, rsmC, rsmD, and rsmH within their genetic contexts, identified their promoter regions, and assessed the impact of both their deletion and overexpression on various Pto DC3000 phenotypes. A novel finding is that rsmA and rsmC are part of an operon with the upstream genes, whereas rsmH seems to be co-transcribed with two downstream genes. We also observed significant variability in expression levels and RpoS dependence among the five rsm paralogs. Thus, despite the extensive repertoire of rsm genes in Pto DC3000, only rsmA , rsmE and rsmH were significantly expressed under all tested conditions (swarming, minimal and T3SS-inducing liquid media). Among these, RsmE and RsmA were corroborated as the most important paralogs at the functional level, whereas RsmH played a minor role in regulating free life and plant-associated phenotypes. Conversely, RsmC and RsmD did not seem to be functional under the conditions tested. [Display omitted] [ABSTRACT FROM AUTHOR]

Published
2024
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2. Exploring the expression and functionality of the rsm sRNAs in Pseudomonas syringae pv. tomato DC3000.

Author

Ferreiro, María-Dolores, Behrmann, Lara Vanessa, Corral, Ana, Nogales, Joaquina, and Gallegos, María-Trinidad

Subjects
PSEUDOMONAS syringae, PROMOTERS (Genetics), TOMATOES, PHENOTYPES, ALGINIC acid
Abstract

The Gac-rsm pathway is a global regulatory network that governs mayor lifestyle and metabolic changes in gamma-proteobacteria. In a previous study, we uncovered the role of CsrA proteins promoting growth and repressing motility, alginate production and virulence in the model phytopathogen Pseudomonas syringae pv. tomato (Pto) DC3000. Here, we focus on the expression and regulation of the rsm regulatory sRNAs, since Pto DC3000 exceptionally has seven variants (rsmX1-5, rsmY and rsmZ). The presented results offer further insights into the functioning of the complex Gac-rsm pathway and the interplay among its components. Overall, rsm expressions reach maximum levels at high cell densities, are unaffected by surface detection, and require GacA for full expression. The rsm levels of expression and GacA-dependence are determined by the sequences found in their −35/-10 promoter regions and GacA binding boxes, respectively. rsmX5 stands out for being the only rsm in Pto DC3000 whose high expression does not require GacA, constituting the main component of the total rsm pool in a gacA mutant. The deletion of rsmY and rsmZ had minor effects on Pto DC3000 motility and virulence phenotypes, indicating that rsmX1-5 can functionally replace them. On the other hand, rsmY or rsmZ overexpression in a gacA mutant did not revert its phenotype. Additionally, a negative feedback regulatory loop in which the CsrA3 protein promotes its own titration by increasing the levels of several rsm RNAs in a GacA-dependent manner has been disclosed as part of this work. [ABSTRACT FROM AUTHOR]

Published
2021
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3. Exploring the expression and functionality of the rsm sRNAs in Pseudomonas syringae pv. tomato DC3000

Author

Ministerio de Ciencia, Innovación y Universidades (España), Ministerio de Educación, Cultura y Deporte (España), German Academic Exchange Service, Ferreiro, María Dolores, Behrmann, L.V., Corral, Ana, Nogales, Joaquina, Gallegos, María Trinidad, Ministerio de Ciencia, Innovación y Universidades (España), Ministerio de Educación, Cultura y Deporte (España), German Academic Exchange Service, Ferreiro, María Dolores, Behrmann, L.V., Corral, Ana, Nogales, Joaquina, and Gallegos, María Trinidad

Abstract

The Gac-rsm pathway is a global regulatory network that governs mayor lifestyle and metabolic changes in gamma-proteobacteria. In a previous study, we uncovered the role of CsrA proteins promoting growth and repressing motility, alginate production and virulence in the model phytopathogen Pseudomonas syringae pv. tomato (Pto) DC3000. Here, we focus on the expression and regulation of the rsm regulatory sRNAs, since Pto DC3000 exceptionally has seven variants (rsmX1-5, rsmY and rsmZ). The presented results offer further insights into the functioning of the complex Gac-rsm pathway and the interplay among its components. Overall, rsm expressions reach maximum levels at high cell densities, are unaffected by surface detection, and require GacA for full expression. The rsm levels of expression and GacA-dependence are determined by the sequences found in their −35/-10 promoter regions and GacA binding boxes, respectively. rsmX5 stands out for being the only rsm in Pto DC3000 whose high expression does not require GacA, constituting the main component of the total rsm pool in a gacA mutant. The deletion of rsmY and rsmZ had minor effects on Pto DC3000 motility and virulence phenotypes, indicating that rsmX1-5 can functionally replace them. On the other hand, rsmY or rsmZ overexpression in a gacA mutant did not revert its phenotype. Additionally, a negative feedback regulatory loop in which the CsrA3 protein promotes its own titration by increasing the levels of several rsm RNAs in a GacA-dependent manner has been disclosed as part of this work.

Published
2021

4. Exploring the expression and functionality of the rsm sRNAs in Pseudomonas syringae pv. tomato DC3000

Author

Lara Vanessa Behrmann, María-Dolores Ferreiro, María-Trinidad Gallegos, Ana Corral, Joaquina Nogales, Ministerio de Ciencia, Innovación y Universidades (España), Ministerio de Educación, Cultura y Deporte (España), and German Academic Exchange Service

Subjects
Genetics, 0303 health sciences, Gac-rsm, RsmA, food and beverages, Pseudomonas syringae, Cell Biology, Biology, 03 medical and health sciences, 0302 clinical medicine, Global regulation, Regulatory RNAs, 030220 oncology & carcinogenesis, sense organs, CsrA, Molecular Biology, 030304 developmental biology
Abstract

The Gac-rsm pathway is a global regulatory network that governs mayor lifestyle and metabolic changes in gamma-proteobacteria. In a previous study, we uncovered the role of CsrA proteins promoting growth and repressing motility, alginate production and virulence in the model phytopathogen Pseudomonas syringae pv. tomato (Pto) DC3000. Here, we focus on the expression and regulation of the rsm regulatory sRNAs, since Pto DC3000 exceptionally has seven variants (rsmX1-5, rsmY and rsmZ). The presented results offer further insights into the functioning of the complex Gac-rsm pathway and the interplay among its components. Overall, rsm expressions reach maximum levels at high cell densities, are unaffected by surface detection, and require GacA for full expression. The rsm levels of expression and GacA-dependence are determined by the sequences found in their −35/-10 promoter regions and GacA binding boxes, respectively. rsmX5 stands out for being the only rsm in Pto DC3000 whose high expression does not require GacA, constituting the main component of the total rsm pool in a gacA mutant. The deletion of rsmY and rsmZ had minor effects on Pto DC3000 motility and virulence phenotypes, indicating that rsmX1-5 can functionally replace them. On the other hand, rsmY or rsmZ overexpression in a gacA mutant did not revert its phenotype. Additionally, a negative feedback regulatory loop in which the CsrA3 protein promotes its own titration by increasing the levels of several rsm RNAs in a GacA-dependent manner has been disclosed as part of this work., This work was supported by the ERDF/Spanish Ministry of Science, Innovation and Universities - State Research Agency under Grants This work was supported by the ERDF/Spanish Ministry of Science, Innovation and Universities - State Research Agency under Grants BIO2014-55075-P and BIO2017-83533-P; Ministerio de Educación, Cultura y Deporte [ECD/1619/2013]; Deutscher Akademischer Austauschdienst (DE) [RISE-Worldwide scholarship]; and BIO2017-83533-P; Ministerio de Educación, Cultura y Deporte [ECD/1619/2013]; Deutscher Akademischer Austauschdienst (DE) [RISE-Worldwide scholarship]

Published
2021

5. A matter of hierarchy: Activation of orfamide production by the posttranscriptional Gac-Rsm cascade of Pseudomonas protegens CHA0 through expression upregulation of the two dedicated transcriptional regulators

Author

Sobrero, Patricio Martín, Muzlera, Andrés, Frescura, Julieta Magali, Jofré, Edgardo, and Valverde, Claudio Fabián

Subjects
Ciencias Biológicas, PSEUDOMONAS PROTEGENS, GAC-RSM, POSTTRANSCRIPTIONAL CONTROL, TRANSCRIPTIONAL REGULATOR, Otras Ciencias Biológicas, ORFAMIDE, CIENCIAS NATURALES Y EXACTAS
Abstract

In this work, we surveyed the genome of P. protegens CHA0 in order to identify novel mRNAs possibly under the control of the Gac-Rsm cascade that might, for their part, serve to elucidate as-yet-unknown functions involved in the biocontrol of plant pathogens and/or in cellular processes required for fitness in natural environments. In view of the experimental evidence from former studies on the Gac-Rsm cascade, we developed a computational screen supported by a combination of sequence, structural, and evolutionary constraints that led to a dataset of 43 potential novel mRNA targets. We then confirmed several mRNA targets experimentally and next focused on two of the respective genes that are physically linked to the orfamide biosynthetic gene cluster and whose predicted open-reading frames resembled cognate LuxR-type transcriptional regulators of cyclic lipopeptide clusters in related pseudomonads. In this report, we demonstrate that in strain CHA0, orfamide production is stringently dependent on a functional Gac-Rsm cascade and that both mRNAs encoding transcriptional regulatory proteins are under direct translational control of the RsmA/E proteins. Our results have thus revealed a hierarchical control over the expression of orfamide biosynthetic genes with the final transcriptional control subordinated to the global Gac-Rsm posttranscriptional regulatory system. Fil: Sobrero, Patricio Martín. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; Argentina Fil: Muzlera, Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; Argentina Fil: Frescura, Julieta Magali. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; Argentina Fil: Jofré, Edgardo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Ciencias Naturales; Argentina Fil: Valverde, Claudio Fabián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; Argentina

Published
2017
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