The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniae
ABSTRACT Eukaryotic-like membrane Ser/Thr protein kinases play a pivotal role in different aspects of bacterial physiology. In contrast to the diversity of their extracellular domains, their cytoplasmic catalytic domains are highly conserved. However, the function of a long juxtamembrane domain (JMD...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2025-05-01
|
| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.03799-24 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850031124596654080 |
|---|---|
| author | Mélisse Hamidi Sathya Narayanan Nagarajan Vaishnavi Ravikumar Virginie Gueguen-Chaignon Cédric Laguri Céline Freton Ivan Mijakovic Jean-Pierre Simorre Stéphanie Ravaud Christophe Grangeasse |
| author_facet | Mélisse Hamidi Sathya Narayanan Nagarajan Vaishnavi Ravikumar Virginie Gueguen-Chaignon Cédric Laguri Céline Freton Ivan Mijakovic Jean-Pierre Simorre Stéphanie Ravaud Christophe Grangeasse |
| author_sort | Mélisse Hamidi |
| collection | DOAJ |
| description | ABSTRACT Eukaryotic-like membrane Ser/Thr protein kinases play a pivotal role in different aspects of bacterial physiology. In contrast to the diversity of their extracellular domains, their cytoplasmic catalytic domains are highly conserved. However, the function of a long juxtamembrane domain (JMD), which connects the catalytic domain to the transmembrane helix, remains elusive. In this study, we investigated the function of the JMD of the Ser/Thr protein kinase StkP in the cell division of Streptococcus pneumoniae. We observed that the deletion of the JMD affected the ability of StkP to phosphorylate some of its endogenous substrates, thereby resulting in significant cell morphogenesis defects. Furthermore, multiple threonine residues were identified as being phosphorylated in the JMD. To investigate the functional significance of these phosphorylation sites, we conducted an integrative analysis, combining structural biology, proteomics, and bacterial cell imaging. Our results revealed that the phosphorylation of the JMD did not perturb the phosphorylation of StkP substrates. However, we observed that it modulated the timing of StkP localization to the division septum and the dynamics of cell constriction. We further demonstrated that phosphorylation of the JMD facilitated the recruitment of several cell division proteins, suggesting that it is required to assemble the division machinery at the division septum. In conclusion, this study demonstrates that the function of the JMD of StkP is modulated by phosphorylation and is critical for the cell division of S. pneumoniae. These observations may serve as a model for understanding the regulatory function of other bacterial Ser/Thr protein kinases.IMPORTANCEHow bacterial serine/threonine protein kinases are activated remains highly debated. In particular, models rely on the observations made with their eukaryotic counterparts, and only a few studies have investigated the molecular activation mechanism of bacterial serine/threonine protein kinases. This is particularly the case with regard to the juxtamembrane domain (JMD), which is proposed to contribute to kinase activation in numerous eukaryotic kinases. This study demonstrates that the juxtamembrane domain is likely not essential for the activation of the serine/threonine protein kinase StkP of S. pneumoniae. Rather, our findings reveal that it is required for cell division, where its phosphorylation affects the assembly of the division machinery at the division septum. These observations allow us to assign a function to the JMD in StkP-mediated regulation of pneumococcal cell division, thereby providing a new avenue for understanding the contribution of membrane serine/threonine protein kinases in the physiology of other bacteria. |
| format | Article |
| id | doaj-art-018e554685bb4e21894067916b6dc439 |
| institution | DOAJ |
| issn | 2150-7511 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj-art-018e554685bb4e21894067916b6dc4392025-08-20T02:59:03ZengAmerican Society for MicrobiologymBio2150-75112025-05-0116510.1128/mbio.03799-24The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniaeMélisse Hamidi0Sathya Narayanan Nagarajan1Vaishnavi Ravikumar2Virginie Gueguen-Chaignon3Cédric Laguri4Céline Freton5Ivan Mijakovic6Jean-Pierre Simorre7Stéphanie Ravaud8Christophe Grangeasse9Molecular Microbiology and Structural Biochemistry, UMR 5086, Université Claude Bernard Lyon 1, CNRS, Lyon, Auvergne-Rhône-Alpes, FranceMolecular Microbiology and Structural Biochemistry, UMR 5086, Université Claude Bernard Lyon 1, CNRS, Lyon, Auvergne-Rhône-Alpes, FranceDepartment of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Västra Götaland County, SwedenProtein Science Facility, CNRS UAR3444, INSERM US8, Université Claude Bernard Lyon 1, Ecole Normale Supérieur de Lyon, Lyon, Auvergne-Rhône-Alpes, FranceInstitut de Biologie Structurale, CEA, CNRS UMR 5075, Université Grenoble Alpes, Grenoble, Auvergne-Rhône-Alpes, FranceMolecular Microbiology and Structural Biochemistry, UMR 5086, Université Claude Bernard Lyon 1, CNRS, Lyon, Auvergne-Rhône-Alpes, FranceDepartment of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Västra Götaland County, SwedenInstitut de Biologie Structurale, CEA, CNRS UMR 5075, Université Grenoble Alpes, Grenoble, Auvergne-Rhône-Alpes, FranceMolecular Microbiology and Structural Biochemistry, UMR 5086, Université Claude Bernard Lyon 1, CNRS, Lyon, Auvergne-Rhône-Alpes, FranceMolecular Microbiology and Structural Biochemistry, UMR 5086, Université Claude Bernard Lyon 1, CNRS, Lyon, Auvergne-Rhône-Alpes, FranceABSTRACT Eukaryotic-like membrane Ser/Thr protein kinases play a pivotal role in different aspects of bacterial physiology. In contrast to the diversity of their extracellular domains, their cytoplasmic catalytic domains are highly conserved. However, the function of a long juxtamembrane domain (JMD), which connects the catalytic domain to the transmembrane helix, remains elusive. In this study, we investigated the function of the JMD of the Ser/Thr protein kinase StkP in the cell division of Streptococcus pneumoniae. We observed that the deletion of the JMD affected the ability of StkP to phosphorylate some of its endogenous substrates, thereby resulting in significant cell morphogenesis defects. Furthermore, multiple threonine residues were identified as being phosphorylated in the JMD. To investigate the functional significance of these phosphorylation sites, we conducted an integrative analysis, combining structural biology, proteomics, and bacterial cell imaging. Our results revealed that the phosphorylation of the JMD did not perturb the phosphorylation of StkP substrates. However, we observed that it modulated the timing of StkP localization to the division septum and the dynamics of cell constriction. We further demonstrated that phosphorylation of the JMD facilitated the recruitment of several cell division proteins, suggesting that it is required to assemble the division machinery at the division septum. In conclusion, this study demonstrates that the function of the JMD of StkP is modulated by phosphorylation and is critical for the cell division of S. pneumoniae. These observations may serve as a model for understanding the regulatory function of other bacterial Ser/Thr protein kinases.IMPORTANCEHow bacterial serine/threonine protein kinases are activated remains highly debated. In particular, models rely on the observations made with their eukaryotic counterparts, and only a few studies have investigated the molecular activation mechanism of bacterial serine/threonine protein kinases. This is particularly the case with regard to the juxtamembrane domain (JMD), which is proposed to contribute to kinase activation in numerous eukaryotic kinases. This study demonstrates that the juxtamembrane domain is likely not essential for the activation of the serine/threonine protein kinase StkP of S. pneumoniae. Rather, our findings reveal that it is required for cell division, where its phosphorylation affects the assembly of the division machinery at the division septum. These observations allow us to assign a function to the JMD in StkP-mediated regulation of pneumococcal cell division, thereby providing a new avenue for understanding the contribution of membrane serine/threonine protein kinases in the physiology of other bacteria.https://journals.asm.org/doi/10.1128/mbio.03799-24StkPStreptococcus pneumoniaeprotein kinasebacterial cell divisionjuxtamembrane domainStkP |
| spellingShingle | Mélisse Hamidi Sathya Narayanan Nagarajan Vaishnavi Ravikumar Virginie Gueguen-Chaignon Cédric Laguri Céline Freton Ivan Mijakovic Jean-Pierre Simorre Stéphanie Ravaud Christophe Grangeasse The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniae mBio StkP Streptococcus pneumoniae protein kinase bacterial cell division juxtamembrane domain StkP |
| title | The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniae |
| title_full | The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniae |
| title_fullStr | The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniae |
| title_full_unstemmed | The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniae |
| title_short | The juxtamembrane domain of StkP is phosphorylated and influences cell division in Streptococcus pneumoniae |
| title_sort | juxtamembrane domain of stkp is phosphorylated and influences cell division in streptococcus pneumoniae |
| topic | StkP Streptococcus pneumoniae protein kinase bacterial cell division juxtamembrane domain StkP |
| url | https://journals.asm.org/doi/10.1128/mbio.03799-24 |
| work_keys_str_mv | AT melissehamidi thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT sathyanarayanannagarajan thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT vaishnaviravikumar thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT virginiegueguenchaignon thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT cedriclaguri thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT celinefreton thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT ivanmijakovic thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT jeanpierresimorre thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT stephanieravaud thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT christophegrangeasse thejuxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT melissehamidi juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT sathyanarayanannagarajan juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT vaishnaviravikumar juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT virginiegueguenchaignon juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT cedriclaguri juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT celinefreton juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT ivanmijakovic juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT jeanpierresimorre juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT stephanieravaud juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae AT christophegrangeasse juxtamembranedomainofstkpisphosphorylatedandinfluencescelldivisioninstreptococcuspneumoniae |