The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and division
ABSTRACT Cell elongation and division are essential aspects of the bacterial life cycle that must be coordinated for viability and replication. The impact of misregulation of these processes is not well understood as these systems are often not amenable to traditional genetic manipulation. Recently,...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-08-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.00631-23 |
| _version_ | 1797730270425645056 |
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| author | Bryan D. Lakey François Alberge Daniel Parrell Elizabeth R. Wright Daniel R. Noguera Timothy J. Donohue |
| author_facet | Bryan D. Lakey François Alberge Daniel Parrell Elizabeth R. Wright Daniel R. Noguera Timothy J. Donohue |
| author_sort | Bryan D. Lakey |
| collection | DOAJ |
| description | ABSTRACT Cell elongation and division are essential aspects of the bacterial life cycle that must be coordinated for viability and replication. The impact of misregulation of these processes is not well understood as these systems are often not amenable to traditional genetic manipulation. Recently, we reported on the CenKR two-component system (TCS) in the Gram-negative bacterium Rhodobacter sphaeroides that is genetically tractable, widely conserved in α-proteobacteria, and directly regulates the expression of components crucial for cell elongation and division, including genes encoding subunit of the Tol-Pal complex. In this work, we show that overexpression of cenK results in cell filamentation and chaining. Using cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET), we generated high-resolution two-dimensional (2D) images and three-dimensional (3D) volumes of the cell envelope and division septum of wild-type cells and a cenK overexpression strain finding that these morphological changes stem from defects in outer membrane (OM) and peptidoglycan (PG) constriction. By monitoring the localization of Pal, PG biosynthesis, and the bacterial cytoskeletal proteins MreB and FtsZ, we developed a model for how increased CenKR activity leads to changes in cell elongation and division. This model predicts that increased CenKR activity decreases the mobility of Pal, delaying OM constriction, and ultimately disrupting the midcell positioning of MreB and FtsZ and interfering with the spatial regulation of PG synthesis and remodeling. IMPORTANCE By coordinating cell elongation and division, bacteria maintain their shape, support critical envelope functions, and orchestrate division. Regulatory and assembly systems have been implicated in these processes in some well-studied Gram-negative bacteria. However, we lack information on these processes and their conservation across the bacterial phylogeny. In R. sphaeroides and other α-proteobacteria, CenKR is an essential two-component system (TCS) that regulates the expression of genes known or predicted to function in cell envelope biosynthesis, elongation, and/or division. Here, we leverage unique features of CenKR to understand how increasing its activity impacts cell elongation/division and use antibiotics to identify how modulating the activity of this TCS leads to changes in cell morphology. Our results provide new insight into how CenKR activity controls the structure and function of the bacterial envelope, the localization of cell elongation and division machinery, and cellular processes in organisms with importance in health, host-microbe interactions, and biotechnology. |
| first_indexed | 2024-03-12T11:41:51Z |
| format | Article |
| id | doaj.art-a0bc4e4db6de4073adc379a5c7bd7d29 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-03-12T11:41:51Z |
| publishDate | 2023-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-a0bc4e4db6de4073adc379a5c7bd7d292023-08-31T15:04:21ZengAmerican Society for MicrobiologymBio2150-75112023-08-0114410.1128/mbio.00631-23The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and divisionBryan D. Lakey0François Alberge1Daniel Parrell2Elizabeth R. Wright3Daniel R. Noguera4Timothy J. Donohue5Wisconsin Energy Institute, Great Lakes Bioenergy Research Center, University of Wisconsin-Madison , Madison, Wisconsin, USAWisconsin Energy Institute, Great Lakes Bioenergy Research Center, University of Wisconsin-Madison , Madison, Wisconsin, USAWisconsin Energy Institute, Great Lakes Bioenergy Research Center, University of Wisconsin-Madison , Madison, Wisconsin, USAWisconsin Energy Institute, Great Lakes Bioenergy Research Center, University of Wisconsin-Madison , Madison, Wisconsin, USAWisconsin Energy Institute, Great Lakes Bioenergy Research Center, University of Wisconsin-Madison , Madison, Wisconsin, USAWisconsin Energy Institute, Great Lakes Bioenergy Research Center, University of Wisconsin-Madison , Madison, Wisconsin, USAABSTRACT Cell elongation and division are essential aspects of the bacterial life cycle that must be coordinated for viability and replication. The impact of misregulation of these processes is not well understood as these systems are often not amenable to traditional genetic manipulation. Recently, we reported on the CenKR two-component system (TCS) in the Gram-negative bacterium Rhodobacter sphaeroides that is genetically tractable, widely conserved in α-proteobacteria, and directly regulates the expression of components crucial for cell elongation and division, including genes encoding subunit of the Tol-Pal complex. In this work, we show that overexpression of cenK results in cell filamentation and chaining. Using cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET), we generated high-resolution two-dimensional (2D) images and three-dimensional (3D) volumes of the cell envelope and division septum of wild-type cells and a cenK overexpression strain finding that these morphological changes stem from defects in outer membrane (OM) and peptidoglycan (PG) constriction. By monitoring the localization of Pal, PG biosynthesis, and the bacterial cytoskeletal proteins MreB and FtsZ, we developed a model for how increased CenKR activity leads to changes in cell elongation and division. This model predicts that increased CenKR activity decreases the mobility of Pal, delaying OM constriction, and ultimately disrupting the midcell positioning of MreB and FtsZ and interfering with the spatial regulation of PG synthesis and remodeling. IMPORTANCE By coordinating cell elongation and division, bacteria maintain their shape, support critical envelope functions, and orchestrate division. Regulatory and assembly systems have been implicated in these processes in some well-studied Gram-negative bacteria. However, we lack information on these processes and their conservation across the bacterial phylogeny. In R. sphaeroides and other α-proteobacteria, CenKR is an essential two-component system (TCS) that regulates the expression of genes known or predicted to function in cell envelope biosynthesis, elongation, and/or division. Here, we leverage unique features of CenKR to understand how increasing its activity impacts cell elongation/division and use antibiotics to identify how modulating the activity of this TCS leads to changes in cell morphology. Our results provide new insight into how CenKR activity controls the structure and function of the bacterial envelope, the localization of cell elongation and division machinery, and cellular processes in organisms with importance in health, host-microbe interactions, and biotechnology.https://journals.asm.org/doi/10.1128/mbio.00631-23Rhodobactercell divisiongram-negative cell envelopetol-paltwo-component systemalphaproteobacteria |
| spellingShingle | Bryan D. Lakey François Alberge Daniel Parrell Elizabeth R. Wright Daniel R. Noguera Timothy J. Donohue The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and division mBio Rhodobacter cell division gram-negative cell envelope tol-pal two-component system alphaproteobacteria |
| title | The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and division |
| title_full | The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and division |
| title_fullStr | The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and division |
| title_full_unstemmed | The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and division |
| title_short | The role of CenKR in the coordination of Rhodobacter sphaeroides cell elongation and division |
| title_sort | role of cenkr in the coordination of rhodobacter sphaeroides cell elongation and division |
| topic | Rhodobacter cell division gram-negative cell envelope tol-pal two-component system alphaproteobacteria |
| url | https://journals.asm.org/doi/10.1128/mbio.00631-23 |
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