Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli

Phasins (PhaP) are proteins normally associated with granules of poly(3-hydroxybutyrate) (PHB), a biodegradable polymer accumulated by many bacteria as a reserve molecule. These proteins enhance growth and polymer production in natural and recombinant PHB producers. It has been shown that the produc...

Descripción completa

Detalles Bibliográficos
Autores principales:	Catone, Mariela Verónica, Pettinari, María Julia
Publicado:	2011
Materias:	Coexpressed E. coli Granule-associated proteins Heat stress Heat-shock Heterologous proteins Misfolded proteins PCR analysis PHB accumulation Poly(3-hydroxybutyrate) Polymer production Protective effects Protein level Recombinant Escherichia coli Reverse transcription Stress condition Stress-related gene Superoxides Biodegradable polymers Escherichia coli Genes Granulation Heat resistance Oxygen Proteins Strain Transcription Biosynthesis bacterial protein chaperone DNA binding protein Escherichia coli protein hydroxybutyric acid PHAP protein, Bacteria poly(3 hydroxybutyric acid) poly-beta-hydroxybutyrate polyester recombinant protein biodegradation coliform bacterium experimental study polymer polymerase chain reaction protein real time recombination temperature tolerance article Azotobacter biosynthesis enzymology gene expression profiling genetics metabolism physiological stress physiology Bacterial Proteins DNA-Binding Proteins Escherichia coli Proteins Gene Expression Profiling Hydroxybutyrates Molecular Chaperones Polyesters Recombinant Proteins Stress, Physiological
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00992240_v77_n18_p6622_Almeida http://hdl.handle.net/20.500.12110/paper_00992240_v77_n18_p6622_Almeida
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_00992240_v77_n18_p6622_Almeida
record_format	dspace
spelling	paper:paper_00992240_v77_n18_p6622_Almeida2023-06-08T15:10:06Z Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli Catone, Mariela Verónica Pettinari, María Julia Coexpressed E. coli Granule-associated proteins Heat stress Heat-shock Heterologous proteins Misfolded proteins PCR analysis PHB accumulation Poly(3-hydroxybutyrate) Polymer production Protective effects Protein level Recombinant Escherichia coli Reverse transcription Stress condition Stress-related gene Superoxides Biodegradable polymers Escherichia coli Genes Granulation Heat resistance Oxygen Proteins Strain Transcription Biosynthesis bacterial protein chaperone DNA binding protein Escherichia coli protein hydroxybutyric acid PHAP protein, Bacteria poly(3 hydroxybutyric acid) poly-beta-hydroxybutyrate polyester recombinant protein biodegradation coliform bacterium experimental study polymer polymerase chain reaction protein real time recombination temperature tolerance article Azotobacter biosynthesis enzymology Escherichia coli gene expression profiling genetics metabolism physiological stress physiology Azotobacter Bacterial Proteins DNA-Binding Proteins Escherichia coli Escherichia coli Proteins Gene Expression Profiling Hydroxybutyrates Molecular Chaperones Polyesters Recombinant Proteins Stress, Physiological Azotobacter Escherichia coli Phasins (PhaP) are proteins normally associated with granules of poly(3-hydroxybutyrate) (PHB), a biodegradable polymer accumulated by many bacteria as a reserve molecule. These proteins enhance growth and polymer production in natural and recombinant PHB producers. It has been shown that the production of PHB causes stress in recombinant Escherichia coli, revealed by an increase in the concentrations of several heat stress proteins. In this work, quantitative reverse transcription (qRT)-PCR analysis was used to study the effect of PHB accumulation, and that of PhaP from Azotobacter sp. strain FA8, on the expression of stress-related genes in PHB-producing E. coli. While PHB accumulation was found to increase the transcription of dnaK and ibpA, the expression of these genes and of groES, groEL, rpoH, dps, and yfiD was reduced, when PhaP was coexpressed, to levels even lower than those detected in the non-PHB-accumulating control. These results demonstrated the protective role of PhaP in PHB-synthesizing E. coli and linked the effects of the protein to the expression of stress-related genes, especially ibpA. The effect of PhaP was also analyzed in non-PHBsynthesizing strains, showing that expression of this heterologous protein has an unexpected protective effect in E. coli, under both normal and stress conditions, resulting in increased growth and higher resistance to both heat shock and superoxide stress by paraquat. In addition, PhaP expression was shown to reduce RpoH protein levels during heat shock, probably by reducing or titrating the levels of misfolded proteins. © 2011, American Society for Microbiology. Fil:Catone, M.V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Pettinari, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2011 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00992240_v77_n18_p6622_Almeida http://hdl.handle.net/20.500.12110/paper_00992240_v77_n18_p6622_Almeida
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Coexpressed E. coli Granule-associated proteins Heat stress Heat-shock Heterologous proteins Misfolded proteins PCR analysis PHB accumulation Poly(3-hydroxybutyrate) Polymer production Protective effects Protein level Recombinant Escherichia coli Reverse transcription Stress condition Stress-related gene Superoxides Biodegradable polymers Escherichia coli Genes Granulation Heat resistance Oxygen Proteins Strain Transcription Biosynthesis bacterial protein chaperone DNA binding protein Escherichia coli protein hydroxybutyric acid PHAP protein, Bacteria poly(3 hydroxybutyric acid) poly-beta-hydroxybutyrate polyester recombinant protein biodegradation coliform bacterium experimental study polymer polymerase chain reaction protein real time recombination temperature tolerance article Azotobacter biosynthesis enzymology Escherichia coli gene expression profiling genetics metabolism physiological stress physiology Azotobacter Bacterial Proteins DNA-Binding Proteins Escherichia coli Escherichia coli Proteins Gene Expression Profiling Hydroxybutyrates Molecular Chaperones Polyesters Recombinant Proteins Stress, Physiological Azotobacter Escherichia coli
spellingShingle	Coexpressed E. coli Granule-associated proteins Heat stress Heat-shock Heterologous proteins Misfolded proteins PCR analysis PHB accumulation Poly(3-hydroxybutyrate) Polymer production Protective effects Protein level Recombinant Escherichia coli Reverse transcription Stress condition Stress-related gene Superoxides Biodegradable polymers Escherichia coli Genes Granulation Heat resistance Oxygen Proteins Strain Transcription Biosynthesis bacterial protein chaperone DNA binding protein Escherichia coli protein hydroxybutyric acid PHAP protein, Bacteria poly(3 hydroxybutyric acid) poly-beta-hydroxybutyrate polyester recombinant protein biodegradation coliform bacterium experimental study polymer polymerase chain reaction protein real time recombination temperature tolerance article Azotobacter biosynthesis enzymology Escherichia coli gene expression profiling genetics metabolism physiological stress physiology Azotobacter Bacterial Proteins DNA-Binding Proteins Escherichia coli Escherichia coli Proteins Gene Expression Profiling Hydroxybutyrates Molecular Chaperones Polyesters Recombinant Proteins Stress, Physiological Azotobacter Escherichia coli Catone, Mariela Verónica Pettinari, María Julia Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli
topic_facet	Coexpressed E. coli Granule-associated proteins Heat stress Heat-shock Heterologous proteins Misfolded proteins PCR analysis PHB accumulation Poly(3-hydroxybutyrate) Polymer production Protective effects Protein level Recombinant Escherichia coli Reverse transcription Stress condition Stress-related gene Superoxides Biodegradable polymers Escherichia coli Genes Granulation Heat resistance Oxygen Proteins Strain Transcription Biosynthesis bacterial protein chaperone DNA binding protein Escherichia coli protein hydroxybutyric acid PHAP protein, Bacteria poly(3 hydroxybutyric acid) poly-beta-hydroxybutyrate polyester recombinant protein biodegradation coliform bacterium experimental study polymer polymerase chain reaction protein real time recombination temperature tolerance article Azotobacter biosynthesis enzymology Escherichia coli gene expression profiling genetics metabolism physiological stress physiology Azotobacter Bacterial Proteins DNA-Binding Proteins Escherichia coli Escherichia coli Proteins Gene Expression Profiling Hydroxybutyrates Molecular Chaperones Polyesters Recombinant Proteins Stress, Physiological Azotobacter Escherichia coli
description	Phasins (PhaP) are proteins normally associated with granules of poly(3-hydroxybutyrate) (PHB), a biodegradable polymer accumulated by many bacteria as a reserve molecule. These proteins enhance growth and polymer production in natural and recombinant PHB producers. It has been shown that the production of PHB causes stress in recombinant Escherichia coli, revealed by an increase in the concentrations of several heat stress proteins. In this work, quantitative reverse transcription (qRT)-PCR analysis was used to study the effect of PHB accumulation, and that of PhaP from Azotobacter sp. strain FA8, on the expression of stress-related genes in PHB-producing E. coli. While PHB accumulation was found to increase the transcription of dnaK and ibpA, the expression of these genes and of groES, groEL, rpoH, dps, and yfiD was reduced, when PhaP was coexpressed, to levels even lower than those detected in the non-PHB-accumulating control. These results demonstrated the protective role of PhaP in PHB-synthesizing E. coli and linked the effects of the protein to the expression of stress-related genes, especially ibpA. The effect of PhaP was also analyzed in non-PHBsynthesizing strains, showing that expression of this heterologous protein has an unexpected protective effect in E. coli, under both normal and stress conditions, resulting in increased growth and higher resistance to both heat shock and superoxide stress by paraquat. In addition, PhaP expression was shown to reduce RpoH protein levels during heat shock, probably by reducing or titrating the levels of misfolded proteins. © 2011, American Society for Microbiology.
author	Catone, Mariela Verónica Pettinari, María Julia
author_facet	Catone, Mariela Verónica Pettinari, María Julia
author_sort	Catone, Mariela Verónica
title	Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli
title_short	Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli
title_full	Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli
title_fullStr	Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli
title_full_unstemmed	Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli
title_sort	unexpected stress-reducing effect of phap, a poly(3-hydroxybutyrate) granule-associated protein, in escherichia coli
publishDate	2011
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00992240_v77_n18_p6622_Almeida http://hdl.handle.net/20.500.12110/paper_00992240_v77_n18_p6622_Almeida
work_keys_str_mv	AT catonemarielaveronica unexpectedstressreducingeffectofphapapoly3hydroxybutyrategranuleassociatedproteininescherichiacoli AT pettinarimariajulia unexpectedstressreducingeffectofphapapoly3hydroxybutyrategranuleassociatedproteininescherichiacoli
_version_	1768543745537474560

Unexpected stress-Reducing effect of PhaP, a poly(3-hydroxybutyrate) granule-associated protein, in Escherichia coli

Ejemplares similares