The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with clover

Rhizobium leguminosarum bv. trifolii is capable of establishing a symbiotic relationship with plants from the genus Trifolium. Previously, a regulatory protein encoded by rosR was identified and characterized in this bacterium. RosR possesses a Cys2-His2-type zinc finger motif and belongs to Ros/Muc...

Full description

Bibliographic Details
Main Authors: Kamila Rachwał, Aleksandra Boguszewska, Joanna Kopcińska, Magdalena Karaś, Marek Tchórzewski, Monika Janczarek
Format: Article
Language:English
Published: Frontiers Media S.A. 2016-08-01
Series:Frontiers in Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01302/full
_version_ 1818289035479613440
author Kamila Rachwał
Aleksandra Boguszewska
Joanna Kopcińska
Magdalena Karaś
Marek Tchórzewski
Monika Janczarek
author_facet Kamila Rachwał
Aleksandra Boguszewska
Joanna Kopcińska
Magdalena Karaś
Marek Tchórzewski
Monika Janczarek
author_sort Kamila Rachwał
collection DOAJ
description Rhizobium leguminosarum bv. trifolii is capable of establishing a symbiotic relationship with plants from the genus Trifolium. Previously, a regulatory protein encoded by rosR was identified and characterized in this bacterium. RosR possesses a Cys2-His2-type zinc finger motif and belongs to Ros/MucR family of rhizobial transcriptional regulators. Transcriptome profiling of the rosR mutant revealed a role of this protein in several cellular processes, including the synthesis of cell-surface components and polysaccharides, motility, and bacterial metabolism. Here, we show that a mutation in rosR resulted in considerable changes in R. leguminosarum bv. trifolii protein profiles. Extracellular, membrane, and periplasmic protein profiles of R. leguminosarum bv. trifolii wild type and the rosR mutant were examined, and proteins with substantially different abundances between these strains were identified. Compared with the wild type, extracellular fraction of the rosR mutant contained greater amounts of several proteins, including Ca2+-binding cadherin-like proteins, a RTX-like protein, autoaggregation protein RapA1, and flagellins FlaA and FlaB. In contrast, several proteins involved in the uptake of various substrates were less abundant in the mutant strain (DppA, BraC, and SfuA). In addition, differences were observed in membrane proteins of the mutant and wild-type strains, which mainly concerned various transport system components. Using atomic force microscopy imaging, we characterized the topography and surface properties of the rosR mutant and wild-type cells. We found that the mutation in rosR gene also affected surface properties of R. leguminosarum bv. trifolii. The mutant cells were significantly more hydrophobic than the wild-type cells, and their outer membrane was three times more permeable to the hydrophobic dye N-phenyl-1-naphthylamine. The mutation of rosR also caused defects in bacterial symbiotic interaction with clover plants. Compared with the wild type, the rosR mutant infected host plant roots much less effectively and its nodule occupation was disturbed. At the ultrastructural level, the most striking differences between the mutant and the wild-type nodules concerned the structure of infection threads, release of bacteria, and bacteroid differentiation. This confirms an essential role of RosR in establishment of successful symbiotic interaction of R. leguminosarum bv. trifolii with clover plants.
first_indexed 2024-12-13T02:05:52Z
format Article
id doaj.art-a9ffd28ca5e14947a5fc63b95cb87a24
institution Directory Open Access Journal
issn 1664-302X
language English
last_indexed 2024-12-13T02:05:52Z
publishDate 2016-08-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Microbiology
spelling doaj.art-a9ffd28ca5e14947a5fc63b95cb87a242022-12-22T00:03:08ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2016-08-01710.3389/fmicb.2016.01302209690The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with cloverKamila Rachwał0Aleksandra Boguszewska1Joanna Kopcińska2Magdalena Karaś3Marek Tchórzewski4Monika Janczarek5Maria Curie-Sklodowska UniversityMaria Curie-Sklodowska UniversityWarsaw University of Life SciencesMaria Curie-Sklodowska UniversityMaria Curie-Sklodowska UniversityMaria Curie-Sklodowska UniversityRhizobium leguminosarum bv. trifolii is capable of establishing a symbiotic relationship with plants from the genus Trifolium. Previously, a regulatory protein encoded by rosR was identified and characterized in this bacterium. RosR possesses a Cys2-His2-type zinc finger motif and belongs to Ros/MucR family of rhizobial transcriptional regulators. Transcriptome profiling of the rosR mutant revealed a role of this protein in several cellular processes, including the synthesis of cell-surface components and polysaccharides, motility, and bacterial metabolism. Here, we show that a mutation in rosR resulted in considerable changes in R. leguminosarum bv. trifolii protein profiles. Extracellular, membrane, and periplasmic protein profiles of R. leguminosarum bv. trifolii wild type and the rosR mutant were examined, and proteins with substantially different abundances between these strains were identified. Compared with the wild type, extracellular fraction of the rosR mutant contained greater amounts of several proteins, including Ca2+-binding cadherin-like proteins, a RTX-like protein, autoaggregation protein RapA1, and flagellins FlaA and FlaB. In contrast, several proteins involved in the uptake of various substrates were less abundant in the mutant strain (DppA, BraC, and SfuA). In addition, differences were observed in membrane proteins of the mutant and wild-type strains, which mainly concerned various transport system components. Using atomic force microscopy imaging, we characterized the topography and surface properties of the rosR mutant and wild-type cells. We found that the mutation in rosR gene also affected surface properties of R. leguminosarum bv. trifolii. The mutant cells were significantly more hydrophobic than the wild-type cells, and their outer membrane was three times more permeable to the hydrophobic dye N-phenyl-1-naphthylamine. The mutation of rosR also caused defects in bacterial symbiotic interaction with clover plants. Compared with the wild type, the rosR mutant infected host plant roots much less effectively and its nodule occupation was disturbed. At the ultrastructural level, the most striking differences between the mutant and the wild-type nodules concerned the structure of infection threads, release of bacteria, and bacteroid differentiation. This confirms an essential role of RosR in establishment of successful symbiotic interaction of R. leguminosarum bv. trifolii with clover plants.http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01302/fullMembrane ProteinsRhizobium leguminosarumSymbiosisCloverextracellular proteinsrosR gene
spellingShingle Kamila Rachwał
Aleksandra Boguszewska
Joanna Kopcińska
Magdalena Karaś
Marek Tchórzewski
Monika Janczarek
The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with clover
Frontiers in Microbiology
Membrane Proteins
Rhizobium leguminosarum
Symbiosis
Clover
extracellular proteins
rosR gene
title The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with clover
title_full The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with clover
title_fullStr The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with clover
title_full_unstemmed The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with clover
title_short The regulatory protein RosR affects Rhizobium leguminosarum bv. trifolii protein profiles, cell surface properties, and symbiosis with clover
title_sort regulatory protein rosr affects rhizobium leguminosarum bv trifolii protein profiles cell surface properties and symbiosis with clover
topic Membrane Proteins
Rhizobium leguminosarum
Symbiosis
Clover
extracellular proteins
rosR gene
url http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.01302/full
work_keys_str_mv AT kamilarachwał theregulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT aleksandraboguszewska theregulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT joannakopcinska theregulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT magdalenakaras theregulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT marektchorzewski theregulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT monikajanczarek theregulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT kamilarachwał regulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT aleksandraboguszewska regulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT joannakopcinska regulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT magdalenakaras regulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT marektchorzewski regulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover
AT monikajanczarek regulatoryproteinrosraffectsrhizobiumleguminosarumbvtrifoliiproteinprofilescellsurfacepropertiesandsymbiosiswithclover