title: Penicillin-binding protein folding is dependent on the PrsA peptidyl-prolyl cis-trans isomerase in Bacillus subtilis.
abstract: Summary The PrsA protein is a membrane-anchored peptidyl-prolyl cis-trans isomerase in Bacillus subtilis and most other Gram-positive bacteria. It catalyses the post-translocational folding of exported proteins and is essential for normal growth of B. subtilis. We studied the mechanism behind this indispensability. We could construct a viable prsA null mutant in the presence of a high concentration of magnesium. Various changes in cell morphology in the absence of PrsA suggested that PrsA is involved in the biosynthesis of the cylindrical lateral wall. Consistently, four penicillin-binding proteins (PBP2a, PBP2b, PBP3 and PBP4) were unstable in the absence of PrsA, while muropeptide analysis revealed a 2% decrease in the peptidoglycan cross-linkage index. Misfolded PBP2a was detected in PrsA-depleted cells, indicating that PrsA is required for the folding of this PBP either directly or indirectly. Furthermore, strongly increased uniform staining of cell wall with a fluorescent vancomycin was observed in the absence of PrsA. We also demonstrated that PrsA is a dimeric or oligomeric protein which is localized at distinct spots organized in a helical pattern along the cell membrane. These results suggest that PrsA is essential for normal growth most probably as PBP folding is dependent on this PPIase.
authors.affiliation: Antimicrobial Resistance Unit, Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare (THL), P.O. Box 30, FI-00271 Helsinki, Finland.
authors.forename: Hanne-Leena
authors.initials: HL
authors.lastname: Hyyryläinen
authors.forename: Bogumila C
authors.initials: BC
authors.lastname: Marciniak
authors.forename: Kathleen
authors.initials: K
authors.lastname: Dahncke
authors.forename: Milla
authors.initials: M
authors.lastname: Pietiäinen
authors.forename: Pascal
authors.initials: P
authors.lastname: Courtin
authors.forename: Marika
authors.initials: M
authors.lastname: Vitikainen
authors.forename: Raili
authors.initials: R
authors.lastname: Seppala
authors.forename: Andreas
authors.initials: A
authors.lastname: Otto
authors.forename: Dörte
authors.initials: D
authors.lastname: Becher
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Chapot-Chartier
authors.forename: Oscar P
authors.initials: OP
authors.lastname: Kuipers
authors.forename: Vesa P
authors.initials: VP
authors.lastname: Kontinen
mesh_terms: D001412:Bacillus subtilis / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry; D002473:Cell Wall / Q000737:chemistry / Q000378:metabolism; D017353:Gene Deletion; D005798:Genes, Bacterial; D020043:Genes, Essential; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D046915:Penicillin-Binding Proteins / Q000378:metabolism*; D010457:Peptidoglycan / Q000032:analysis; D019696:Peptidylprolyl Isomerase / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D055503:Protein Multimerization
chemical_list: D001426:Bacterial Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan; C082160:prsA protein, bacteria; D019696:Peptidylprolyl Isomerase; D008274:Magnesium