Structural and functional investigation of the Small Ribosomal Subunit Biogenesis GTP ase A (RsgA) from Pseudomonas aeruginosa
| Autor: | Francesco Malatesta, Daniele Santorelli, Adele Di Matteo, S. Rocchio, Carlo Travaglini-Allocatelli, Mimma Franceschini, Serena Rinaldo, Francesco Imperi, Luca Federici |
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| Přispěvatelé: | Rocchio, S., Santorelli, D., Rinaldo, S., Franceschini, M., Malatesta, F., Imperi, F., Federici, L., Travaglini-Allocatelli, C., Di Matteo, A. |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
crystal structure GTP' Protein Conformation Protein subunit Molecular Conformation Protein Data Bank (RCSB PDB) RNA-binding protein GTPase Guanosine Diphosphate Biochemistry GTP Phosphohydrolases Pseudomonas aeruginosa 03 medical and health sciences binding kinetic 0302 clinical medicine Protein structure Drug Resistance Bacterial binding kinetics Escherichia coli 30S RsgA Molecular Biology ribosome assembly factors Binding Sites Chemistry Bacterial Infections Cell Biology Ribosome Subunits Small Cell biology Kinetics 030104 developmental biology ribosome assembly factor 030220 oncology & carcinogenesis Biogenesis Protein Binding |
| Zdroj: | The FEBS journal 286 (2019): 4245–4260. doi:10.1111/febs.14959 info:cnr-pdr/source/autori:Rocchio, Serena; Santorelli, Daniele; Rinaldo, Serena; Franceschini, Mimma; Malatesta, Francesco; Imperi, Francesco; Federici, Luca; Travaglini-Allocatelli, Carlo; Di Matteo, Adele/titolo:Structural and functional investigation of the Small Ribosomal Subunit Biogenesis GTPase A (RsgA) from Pseudomonas aeruginosa/doi:10.1111%2Ffebs.14959/rivista:The FEBS journal (Print)/anno:2019/pagina_da:4245/pagina_a:4260/intervallo_pagine:4245–4260/volume:286 |
| ISSN: | 1742-4658 1742-464X |
| DOI: | 10.1111/febs.14959 |
| Popis: | The Small Ribosomal Subunit Biogenesis GTPase A (RsgA) is a bacterial assembly factor involved in the late stages of the 30S subunit maturation. It is a multidomain GTPase in which the central circularly permutated GTPase domain is flanked by an OB domain and a Zn-binding domain. All three domains participate in the interaction with the 30S particle thus ensuring an efficient coupling between catalytic activity and biological function. Invivo studies suggested the relevance of rsgA in bacterial growth and cellular viability, but other pleiotropic roles of RsgA are also emerging. Here, we report the 3D structure of RsgA from Pseudomonasaeruginosa (PaRsgA) in the GDP-bound form. We also report a biophysical and biochemical characterization of the protein in both the GDP-bound and its nucleotide-free form. In particular, we report a kinetic analysis of the RsgA binding to GTP and GDP. We found that PaRsgA is able to bind both nucleotides with submicromolar affinity. The higher affinity towards GDP (KD=0.011μm) with respect to GTP (KD=0.16μm) is mainly ascribed to a smaller GDP dissociation rate. Our results confirm that PaRsgA, like most other GTPases, has a weak intrinsic enzymatic activity (kCAT=0.058min−1). Finally, the biological role of RsgA in P.aeruginosa was investigated, allowing us to conclude that rsgA is dispensable for P.aeruginosa growth but important for drug resistance and virulence in an animal infection model. Databases: Coordinates and structure factors for the protein structure described in this manuscript have been deposited in the Protein Data Bank (https://www.rcsb.org) with the accession code 6H4D. |
| Databáze: | OpenAIRE |
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