The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilis
Regulated ATP-dependent proteolysis is a common feature of developmental processes and plays also a crucial role during environmental perturbations such as stress and starvation. The Bacillus subtilis MgsR regulator controls a subregulon within the stress- and stationary phase σB regulon. After etha...
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Frontiers Media S.A.
2020-05-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2020.00900/full |
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| author | Lars Lilge Alexander Reder Frank Tippmann Friedrich Morgenroth Janice Grohmann Dörte Becher Katharina Riedel Uwe Völker Michael Hecker Michael Hecker Ulf Gerth |
| author_facet | Lars Lilge Alexander Reder Frank Tippmann Friedrich Morgenroth Janice Grohmann Dörte Becher Katharina Riedel Uwe Völker Michael Hecker Michael Hecker Ulf Gerth |
| author_sort | Lars Lilge |
| collection | DOAJ |
| description | Regulated ATP-dependent proteolysis is a common feature of developmental processes and plays also a crucial role during environmental perturbations such as stress and starvation. The Bacillus subtilis MgsR regulator controls a subregulon within the stress- and stationary phase σB regulon. After ethanol exposition and a short time-window of activity, MgsR is ClpXP-dependently degraded with a half-life of approximately 6 min. Surprisingly, a protein interaction analysis with MgsR revealed an association with the McsB arginine kinase and an in vivo degradation assay confirmed a strong impact of McsB on MgsR degradation. In vitro phosphorylation experiments with arginine (R) by lysine (K) substitutions in McsB and its activator McsA unraveled all R residues, which are essentially needed for the arginine kinase reaction. Subsequently, site directed mutagenesis of the MgsR substrate was used to substitute all arginine residues with glutamate (R-E) to mimic arginine phosphorylation and to test their influence on MgsR degradation in vivo. It turned out, that especially the R33E and R94/95E residues (RRPI motif), the latter are adjacently located to the two redox-sensitive cysteines in a 3D model, have the potential to accelerate MgsR degradation. These results imply that selective arginine phosphorylation may have favorable effects for Clp dependent degradation of short-living regulatory proteins. We speculate that in addition to its kinase activity and adaptor function for the ClpC ATPase, McsB might also serve as a proteolytic adaptor for the ClpX ATPase in the degradation mechanism of MgsR. |
| first_indexed | 2024-12-22T20:48:49Z |
| format | Article |
| id | doaj.art-f1d90af889a64a239cb65e81f5e11733 |
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| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-22T20:48:49Z |
| publishDate | 2020-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-f1d90af889a64a239cb65e81f5e117332022-12-21T18:13:09ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2020-05-011110.3389/fmicb.2020.00900519873The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilisLars Lilge0Alexander Reder1Frank Tippmann2Friedrich Morgenroth3Janice Grohmann4Dörte Becher5Katharina Riedel6Uwe Völker7Michael Hecker8Michael Hecker9Ulf Gerth10Institute of Microbiology, University of Greifswald, Greifswald, GermanyInterfaculty Institute for Genetics and Functional Genomics, University of Greifswald, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInterfaculty Institute for Genetics and Functional Genomics, University of Greifswald, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInstitute of Marine Biotechnology, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyRegulated ATP-dependent proteolysis is a common feature of developmental processes and plays also a crucial role during environmental perturbations such as stress and starvation. The Bacillus subtilis MgsR regulator controls a subregulon within the stress- and stationary phase σB regulon. After ethanol exposition and a short time-window of activity, MgsR is ClpXP-dependently degraded with a half-life of approximately 6 min. Surprisingly, a protein interaction analysis with MgsR revealed an association with the McsB arginine kinase and an in vivo degradation assay confirmed a strong impact of McsB on MgsR degradation. In vitro phosphorylation experiments with arginine (R) by lysine (K) substitutions in McsB and its activator McsA unraveled all R residues, which are essentially needed for the arginine kinase reaction. Subsequently, site directed mutagenesis of the MgsR substrate was used to substitute all arginine residues with glutamate (R-E) to mimic arginine phosphorylation and to test their influence on MgsR degradation in vivo. It turned out, that especially the R33E and R94/95E residues (RRPI motif), the latter are adjacently located to the two redox-sensitive cysteines in a 3D model, have the potential to accelerate MgsR degradation. These results imply that selective arginine phosphorylation may have favorable effects for Clp dependent degradation of short-living regulatory proteins. We speculate that in addition to its kinase activity and adaptor function for the ClpC ATPase, McsB might also serve as a proteolytic adaptor for the ClpX ATPase in the degradation mechanism of MgsR.https://www.frontiersin.org/article/10.3389/fmicb.2020.00900/fullClp proteolysisMcsB arginine kinaseMgsR degradationarginine phosphorylationMgsR activity |
| spellingShingle | Lars Lilge Alexander Reder Frank Tippmann Friedrich Morgenroth Janice Grohmann Dörte Becher Katharina Riedel Uwe Völker Michael Hecker Michael Hecker Ulf Gerth The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilis Frontiers in Microbiology Clp proteolysis McsB arginine kinase MgsR degradation arginine phosphorylation MgsR activity |
| title | The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilis |
| title_full | The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilis |
| title_fullStr | The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilis |
| title_full_unstemmed | The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilis |
| title_short | The Involvement of the McsB Arginine Kinase in Clp-Dependent Degradation of the MgsR Regulator in Bacillus subtilis |
| title_sort | involvement of the mcsb arginine kinase in clp dependent degradation of the mgsr regulator in bacillus subtilis |
| topic | Clp proteolysis McsB arginine kinase MgsR degradation arginine phosphorylation MgsR activity |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2020.00900/full |
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