scholarly journals Harnessing β-Lactam Antibiotics for Illumination of the Activity of Penicillin-Binding Proteins in Bacillus subtilis

2020 ◽  
Vol 15 (5) ◽  
pp. 1242-1251 ◽  
Author(s):  
Shabnam Sharifzadeh ◽  
Felix Dempwolff ◽  
Daniel B. Kearns ◽  
Erin E. Carlson
Keyword(s):  
Bacillus Subtilis ◽  
Binding Proteins ◽  
Penicillin Binding Proteins

Dynamic localization of penicillin-binding proteins during spore development in Bacillus subtilis

Microbiology ◽  
2005 ◽  
Vol 151 (3) ◽  
pp. 999-1012 ◽  
Author(s):  
Dirk-Jan Scheffers
Keyword(s):  
Bacillus Subtilis ◽  
Green Fluorescent Protein ◽  
Binding Proteins ◽  
Cytoplasmic Membrane ◽  
Fluorescent Protein ◽  
Spore Formation ◽  
The Other ◽  
Spore Development ◽  
Penicillin Binding Proteins ◽  
Green Fluorescent

During Bacillus subtilis spore formation, many membrane proteins that function in spore development localize to the prespore septum and, subsequently, to the outer prespore membrane. Recently, it was shown that the cell-division-specific penicillin-binding proteins (PBPs) 1 and 2b localize to the asymmetric prespore septum. Here, the author studied the localization of other PBPs, fused to green fluorescent protein (GFP), during spore formation. Fusions to PBPs 4, 2c, 2d, 2a, 3, H, 4b, 5, 4a, 4* and X were expressed during vegetative growth, and their localization was monitored during sporulation. Of these PBPs, 2c, 2d, 4b and 4* have been implicated as having a function in sporulation. It was found that PBP2c, 2d and X changed their localization, while the other PBPs tested were not affected. The putative endopeptidase PbpX appears to spiral out in a pattern that resembles FtsZ redistribution during sporulation, but a pbpX knockout strain had no distinguishable phenotype. PBP2c and 2d localize to the prespore septum and follow the membrane during engulfment, and so are redistributed to the prespore membrane. A similar pattern was observed when GFP–PBP2c was expressed in the mother cell from a sporulation-specific promoter. This work shows that various PBPs known to function during sporulation are redistributed from the cytoplasmic membrane to the prespore.

scholarly journals Harnessing β-Lactam Antibiotics for Illumination of the Activity of Penicillin-Binding Proteins in Bacillus subtilis

2019 ◽  
Author(s):  
Shabnam Sharifzadeh ◽  
Felix Dempwolff ◽  
Daniel B. Kearns ◽  
Erin E. Carlson
Keyword(s):  
Bacillus Subtilis ◽  
Binding Proteins ◽  
Single Cells ◽  
Growth Cycle ◽  
Chemical Probes ◽  
Selective Inhibitors ◽  
Penicillin Binding Proteins ◽  
Peptidoglycan Biosynthesis ◽  
Division Site ◽  
Almost All

ABSTRACTSelective chemical probes enable individual investigation of penicillin-binding proteins (PBPs) and provide critical information about their enzymatic activity with spatial and temporal resolution. To identify scaffolds for novel probes to study peptidoglycan biosynthesis in Bacillus subtilis, we evaluated the PBP inhibition profiles of 21 β-lactam antibiotics from different structural subclasses using a fluorescence-based assay. Most compounds readily labeled PBP1, PBP2a, PBP2b or PBP4. Almost all penicillin scaffolds were co-selective for all or combinations of PBP2a, 2b and 4. Cephalosporins, on the other hand, possessed the lowest IC50 values for PBP1 alone or along with PBP4 (ceftriaxone, cefoxitin), 2b (cefotaxime) or 2a, 2b and 4 (cephalothin). Overall, five selective inhibitors for PBP1 (aztreonam, faropenem, piperacillin, cefuroxime and cefsulodin), one selective inhibitor for PBP5 (6-aminopenicillanic acid) and various co-selective inhibitors for other PBPs in B. subtilis were discovered. Surprisingly, carbapenems strongly inhibited PBP3, formerly shown to have low affinity for β-lactams and speculated to be involved in resistance in B. subtilis. To investigate the specific roles of PBP3, we developed activity-based probes based on the meropenem core and utilized them to monitor the activity of PBP3 in living cells. We showed that PBP3 activity localizes as patches in single cells and concentrates as a ring at the septum and the division site during the cell growth cycle. Our activity-based approach enabled spatial resolution of the transpeptidation activity of individual PBPs in this model microorganism, which was not possible with previous chemical and biological approaches.

scholarly journals Several distinct localization patterns for penicillin-binding proteins in Bacillus subtilis

2003 ◽  
Vol 51 (3) ◽  
pp. 749-764 ◽  
Author(s):  
Dirk-Jan Scheffers ◽  
Laura J. F. Jones ◽  
Jeffery Errington
Keyword(s):  
Bacillus Subtilis ◽  
Binding Proteins ◽  
Penicillin Binding Proteins

scholarly journals Rod Shape Determination by the Bacillus subtilis Class B Penicillin-Binding Proteins Encoded by pbpA and pbpH

2003 ◽  
Vol 185 (16) ◽  
pp. 4717-4726 ◽  
Author(s):  
Yuping Wei ◽  
Teresa Havasy ◽  
Derrell C. McPherson ◽  
David L. Popham
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Binding Proteins ◽  
Structural Integrity ◽  
Sequence Similarity ◽  
Inducible Promoter ◽  
Penicillin Binding Proteins ◽  
Peptidoglycan Cell Wall ◽  
Class B ◽  
Shape Determination

ABSTRACT The peptidoglycan cell wall determines the shape and structural integrity of a bacterial cell. Class B penicillin-binding proteins (PBPs) carry a transpeptidase activity that cross-links peptidoglycan strands via their peptide side chains, and some of these proteins are directly involved in cell shape determination. No Bacillus subtilis PBP with a clear role in rod shape maintenance has been identified. However, previous studies showed that during outgrowth of pbpA mutant spores, the cells grew in an ovoid shape for several hours before they recovered and took on a normal rod shape. It was postulated that another PBP, expressed later during outgrowth, was able to compensate for the lack of the pbpA product, PBP2a, and to guide the formation of a rod shape. The B. subtilis pbpH (ykuA) gene product is predicted to be a class B PBP with greatest sequence similarity to PBP2a. We found that a pbpH-lacZ fusion was expressed at very low levels in early log phase and increased in late log phase. A pbpH null mutant was indistinguishable from the wild-type, but a pbpA pbpH double mutant was nonviable. When pbpH was placed under the control of an inducible promoter in a pbpA mutant, viability was dependent on pbpH expression. Growth of this strain in the absence of inducer resulted in conversion of the cells from rods to ovoid/round shapes and lysis. We conclude that PBP2a and PbpH play redundant roles in formation of a rod-shaped peptidoglycan cell wall.

scholarly journals Penicillin-binding proteins in Bacillus subtilis mutants.

1982 ◽  
Vol 21 (6) ◽  
pp. 979-983 ◽  
Author(s):  
G Kleppe ◽  
W Yu ◽  
J L Strominger
Keyword(s):  
Bacillus Subtilis ◽  
Binding Proteins ◽  
Penicillin Binding Proteins

scholarly journals A Mother Cell-Specific Class B Penicillin-Binding Protein, PBP4b, in Bacillus subtilis

2004 ◽  
Vol 186 (1) ◽  
pp. 258-261 ◽  
Author(s):  
Yuping Wei ◽  
Derrell C. McPherson ◽  
David L. Popham
Keyword(s):  
Bacillus Subtilis ◽  
Rna Polymerase ◽  
Mother Cell ◽  
Binding Proteins ◽  
Binding Protein ◽  
Specific Class ◽  
Penicillin Binding Protein ◽  
Penicillin Binding Proteins ◽  
Peptidoglycan Structure ◽  
Class B

ABSTRACT The Bacillus subtilis genome encodes 16 penicillin-binding proteins (PBPs), some of which are involved in synthesis of the spore peptidoglycan. The pbpI (yrrR) gene encodes a class B PBP, PBP4b, and is transcribed in the mother cell by RNA polymerase containing σE. Loss of PBP4b, alone and in combination with other sporulation-specific PBPs, had no effect on spore peptidoglycan structure.

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