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Genetic analysis of the septal peptidoglycan synthase FtsWI complex supports a conserved activation mechanism for SEDS-bPBP complexes
- Source :
- PLoS Genetics, Vol 17, Iss 4, p e1009366 (2021), PLoS Genetics
- Publication Year :
- 2021
- Publisher :
- Public Library of Science (PLoS), 2021.
-
Abstract
- SEDS family peptidoglycan (PG) glycosyltransferases, RodA and FtsW, require their cognate transpeptidases PBP2 and FtsI (class B penicillin binding proteins) to synthesize PG along the cell cylinder and at the septum, respectively. The activities of these SEDS-bPBPs complexes are tightly regulated to ensure proper cell elongation and division. In Escherichia coli FtsN switches FtsA and FtsQLB to the active forms that synergize to stimulate FtsWI, but the exact mechanism is not well understood. Previously, we isolated an activation mutation in ftsW (M269I) that allows cell division with reduced FtsN function. To try to understand the basis for activation we isolated additional substitutions at this position and found that only the original substitution produced an active mutant whereas drastic changes resulted in an inactive mutant. In another approach we isolated suppressors of an inactive FtsL mutant and obtained FtsWE289G and FtsIK211I and found they bypassed FtsN. Epistatic analysis of these mutations and others confirmed that the FtsN-triggered activation signal goes from FtsQLB to FtsI to FtsW. Mapping these mutations, as well as others affecting the activity of FtsWI, on the RodA-PBP2 structure revealed they are located at the interaction interface between the extracellular loop 4 (ECL4) of FtsW and the pedestal domain of FtsI (PBP3). This supports a model in which the interaction between the ECL4 of SEDS proteins and the pedestal domain of their cognate bPBPs plays a critical role in the activation mechanism.<br />Author summary Bacterial cell division requires the synthesis of septal peptidoglycan by the widely conserved SEDS-bPBP protein complex FtsWI, but how the complex is activated during cell division is still poorly understood. Previous studies suggested that FtsN initiates a signaling cascade in the periplasm to activate FtsWI. Here we isolated and characterized activated FtsW and FtsI mutants and confirmed that the signaling cascade for FtsW activation goes from FtsN to FtsQLB to FtsI and then to FtsW. The residues corresponding to mutations affecting FtsWI activation are clustered to a small region of the interaction interface between the pedestal domain of FtsI and the extracellular loop 4 of FtsW, suggesting that this interaction mediates activation of FtsW. This is strikingly similar to the proposed activation mechanism for the RodA-PBP2 complex, another SEDS-bPBP complex required for cell elongation. Thus, the two homologous SEDS-bPBP complexes are activated similarly by completely unrelated activators that modulate the interaction interface between the SEDS proteins and the bPBPs.
- Subjects :
- Models, Molecular
Cancer Research
Penicillin binding proteins
Cell division
Protein Conformation
Mutant
QH426-470
medicine.disease_cause
Biochemistry
chemistry.chemical_compound
Cell Signaling
Antibiotics
Medicine and Health Sciences
Cell Cycle and Cell Division
Genetics (clinical)
Mutation
Antimicrobials
Organic Compounds
Escherichia coli Proteins
Monosaccharides
Drugs
Signaling Cascades
Cell biology
Chemistry
Cell Processes
Physical Sciences
Hyperexpression Techniques
Research Article
Signal Transduction
Substitution Mutation
Carbohydrates
Peptidoglycan
Library Screening
Biology
Research and Analysis Methods
Microbiology
Protein–protein interaction
Bacterial Proteins
Microbial Control
Gene Expression and Vector Techniques
Genetics
medicine
Penicillin-Binding Proteins
Protein Interactions
Molecular Biology Techniques
Molecular Biology
Ecology, Evolution, Behavior and Systematics
Pharmacology
Molecular Biology Assays and Analysis Techniques
Point mutation
Organic Chemistry
Chemical Compounds
Membrane Proteins
Biology and Life Sciences
Proteins
Periplasmic space
Cell Biology
Arabinose
chemistry
Multiprotein Complexes
Peptidyl Transferases
Peptidoglycan Glycosyltransferase
FtsA
Function (biology)
Subjects
Details
- ISSN :
- 15537404
- Volume :
- 17
- Database :
- OpenAIRE
- Journal :
- PLOS Genetics
- Accession number :
- edsair.doi.dedup.....722c2909914653771216c0daba3f22f7