CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus
CCA-adding enzymes are highly specific RNA polymerases that add and maintain the sequence C-C-A at tRNA 3‘-ends. Recently, we could reveal that cold adaptation of such a polymerase is not only achieved at the expense of enzyme stability, but also at the cost of polymerization fidelity. Enzymes from...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-01-01
|
| Series: | Computational and Structural Biotechnology Journal |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037021004402 |
| _version_ | 1818871960926420992 |
|---|---|
| author | Raphaël de Wijn Kévin Rollet Felix G.M. Ernst Karolin Wellner Heike Betat Mario Mörl Claude Sauter |
| author_facet | Raphaël de Wijn Kévin Rollet Felix G.M. Ernst Karolin Wellner Heike Betat Mario Mörl Claude Sauter |
| author_sort | Raphaël de Wijn |
| collection | DOAJ |
| description | CCA-adding enzymes are highly specific RNA polymerases that add and maintain the sequence C-C-A at tRNA 3‘-ends. Recently, we could reveal that cold adaptation of such a polymerase is not only achieved at the expense of enzyme stability, but also at the cost of polymerization fidelity. Enzymes from psychrophilic organisms usually show an increased structural flexibility to enable catalysis at low temperatures. Here, polymerases face a dilemma, as there is a discrepancy between the need for a tightly controlled flexibility during polymerization and an increased flexibility as strategy for cold adaptation. Based on structural and biochemical analyses, we contribute to clarify the cold adaptation strategy of the psychrophilic CCA-adding enzyme from Planococcus halocryophilus, a gram-positive bacterium thriving in the arctic permafrost at low temperatures down to −15 °C. A comparison with the closely related enzyme from the thermophilic bacterium Geobacillus stearothermophilus reveals several features of cold adaptation - a significantly reduced amount of alpha-helical elements in the C-terminal tRNA-binding region and a structural adaptation in one of the highly conserved catalytic core motifs located in the N-terminal catalytic core of the enzyme. |
| first_indexed | 2024-12-19T12:31:13Z |
| format | Article |
| id | doaj.art-483aeb0567bc4cec89987f2a9e1502ba |
| institution | Directory Open Access Journal |
| issn | 2001-0370 |
| language | English |
| last_indexed | 2024-12-19T12:31:13Z |
| publishDate | 2021-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Computational and Structural Biotechnology Journal |
| spelling | doaj.art-483aeb0567bc4cec89987f2a9e1502ba2022-12-21T20:21:23ZengElsevierComputational and Structural Biotechnology Journal2001-03702021-01-011958455855CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilusRaphaël de Wijn0Kévin Rollet1Felix G.M. Ernst2Karolin Wellner3Heike Betat4Mario Mörl5Claude Sauter6Architecture et Réactivité de l’ARN, Université de Strasbourg, CNRS, IBMC, 67084 Strasbourg, FranceArchitecture et Réactivité de l’ARN, Université de Strasbourg, CNRS, IBMC, 67084 Strasbourg, France; Institute for Biochemistry, Leipzig University, Brüderstr. 34, 04103 Leipzig, GermanyInstitute for Biochemistry, Leipzig University, Brüderstr. 34, 04103 Leipzig, GermanyInstitute for Biochemistry, Leipzig University, Brüderstr. 34, 04103 Leipzig, GermanyInstitute for Biochemistry, Leipzig University, Brüderstr. 34, 04103 Leipzig, GermanyInstitute for Biochemistry, Leipzig University, Brüderstr. 34, 04103 Leipzig, Germany; Corresponding authors.Architecture et Réactivité de l’ARN, Université de Strasbourg, CNRS, IBMC, 67084 Strasbourg, France; Corresponding authors.CCA-adding enzymes are highly specific RNA polymerases that add and maintain the sequence C-C-A at tRNA 3‘-ends. Recently, we could reveal that cold adaptation of such a polymerase is not only achieved at the expense of enzyme stability, but also at the cost of polymerization fidelity. Enzymes from psychrophilic organisms usually show an increased structural flexibility to enable catalysis at low temperatures. Here, polymerases face a dilemma, as there is a discrepancy between the need for a tightly controlled flexibility during polymerization and an increased flexibility as strategy for cold adaptation. Based on structural and biochemical analyses, we contribute to clarify the cold adaptation strategy of the psychrophilic CCA-adding enzyme from Planococcus halocryophilus, a gram-positive bacterium thriving in the arctic permafrost at low temperatures down to −15 °C. A comparison with the closely related enzyme from the thermophilic bacterium Geobacillus stearothermophilus reveals several features of cold adaptation - a significantly reduced amount of alpha-helical elements in the C-terminal tRNA-binding region and a structural adaptation in one of the highly conserved catalytic core motifs located in the N-terminal catalytic core of the enzyme.http://www.sciencedirect.com/science/article/pii/S2001037021004402CCA-adding enzymetRNACold adaptationPsychrophilic proteinX-ray crystallographySAXS |
| spellingShingle | Raphaël de Wijn Kévin Rollet Felix G.M. Ernst Karolin Wellner Heike Betat Mario Mörl Claude Sauter CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus Computational and Structural Biotechnology Journal CCA-adding enzyme tRNA Cold adaptation Psychrophilic protein X-ray crystallography SAXS |
| title | CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus |
| title_full | CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus |
| title_fullStr | CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus |
| title_full_unstemmed | CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus |
| title_short | CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus |
| title_sort | cca addition in the cold structural characterization of the psychrophilic cca adding enzyme from the permafrost bacterium planococcus halocryophilus |
| topic | CCA-adding enzyme tRNA Cold adaptation Psychrophilic protein X-ray crystallography SAXS |
| url | http://www.sciencedirect.com/science/article/pii/S2001037021004402 |
| work_keys_str_mv | AT raphaeldewijn ccaadditioninthecoldstructuralcharacterizationofthepsychrophilicccaaddingenzymefromthepermafrostbacteriumplanococcushalocryophilus AT kevinrollet ccaadditioninthecoldstructuralcharacterizationofthepsychrophilicccaaddingenzymefromthepermafrostbacteriumplanococcushalocryophilus AT felixgmernst ccaadditioninthecoldstructuralcharacterizationofthepsychrophilicccaaddingenzymefromthepermafrostbacteriumplanococcushalocryophilus AT karolinwellner ccaadditioninthecoldstructuralcharacterizationofthepsychrophilicccaaddingenzymefromthepermafrostbacteriumplanococcushalocryophilus AT heikebetat ccaadditioninthecoldstructuralcharacterizationofthepsychrophilicccaaddingenzymefromthepermafrostbacteriumplanococcushalocryophilus AT mariomorl ccaadditioninthecoldstructuralcharacterizationofthepsychrophilicccaaddingenzymefromthepermafrostbacteriumplanococcushalocryophilus AT claudesauter ccaadditioninthecoldstructuralcharacterizationofthepsychrophilicccaaddingenzymefromthepermafrostbacteriumplanococcushalocryophilus |