TY - JOUR
T1 - Peptidoglycan precursor synthesis along the sidewall of pole-growing mycobacteria
AU - Pohane, Amol A
AU - Swarts, Benjamin
N1 - Funding Information:
We are grateful to Dr. Kasia Baranowski, Dr. Eric Rubin and Dr. Hesper Rego for sharingL,D-trans-peptidase mutants and for providing critical feedback. We thank Dr. Suzanne Walker and Dr. Kaitlin Schaefer for the S. aureus pbp4 expression construct and for helpful technical advice; Dr. Steven Sandler, Dr. Peter Chien, Dr. James Chambers and Dr. Amy Burnside for microscopy and flow cytometry guidance; Ms. Sylvia Rivera for technical assistance; Dr. Krista Gile for guidance on statistical analysis. We also acknowledge Dr. Chris Sassetti for the MurJ depletion strain, Dr. Graham Hatfull for the ddlAts mutant, Dr. William Jacobs for ΔRD1 ΔpanCD M. tuberculosis and Dr. Yves Brun, Dr. Erkin Kuru and Dr. Michael VanNieuwenhze for the initial supply of the alkDADA (EDA-DA) probe.Funder Grant reference number Author National Institutes of Health New Innovator Award DP2 AI138238 M Sloan Siegrist National Science Foundation CAREER 1654408 Benjamin M Swarts Simons Foundation Life Sciences Research Foundation Fellowship Hoong Chuin Lim Research Corporation for Science Advancement Cottrell College Science Award 22525 Benjamin M Swarts National Institutes of Health U01CA221230 M Sloan Siegrist National Institutes of Health Training Grant T32 GM008515 Emily S Melzer The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Publisher Copyright:
© García-Heredia et al.
PY - 2018
Y1 - 2018
N2 - Rod-shaped mycobacteria expand from their poles, yet d-amino acid probes label cell wall peptidoglycan in this genus at both the poles and sidewall. We sought to clarify the metabolic fates of these probes. Monopeptide incorporation was decreased by antibiotics that block peptidoglycan synthesis or l,d-transpeptidation and in an l,d-transpeptidase mutant. Dipeptides complemented defects in d-alanine synthesis or ligation and were present in lipid-linked peptidoglycan precursors. Characterizing probe uptake pathways allowed us to localize peptidoglycan metabolism with precision: monopeptide-marked l,d-transpeptidase remodeling and dipeptide-marked synthesis were coincident with mycomembrane metabolism at the poles, septum and sidewall. Fluorescent pencillin-marked d,d-transpeptidation around the cell perimeter further suggested that the mycobacterial sidewall is a site of cell wall assembly. While polar peptidoglycan synthesis was associated with cell elongation, sidewall synthesis responded to cell wall damage. Peptidoglycan editing along the sidewall may support cell wall robustness in pole-growing mycobacteria.
AB - Rod-shaped mycobacteria expand from their poles, yet d-amino acid probes label cell wall peptidoglycan in this genus at both the poles and sidewall. We sought to clarify the metabolic fates of these probes. Monopeptide incorporation was decreased by antibiotics that block peptidoglycan synthesis or l,d-transpeptidation and in an l,d-transpeptidase mutant. Dipeptides complemented defects in d-alanine synthesis or ligation and were present in lipid-linked peptidoglycan precursors. Characterizing probe uptake pathways allowed us to localize peptidoglycan metabolism with precision: monopeptide-marked l,d-transpeptidase remodeling and dipeptide-marked synthesis were coincident with mycomembrane metabolism at the poles, septum and sidewall. Fluorescent pencillin-marked d,d-transpeptidation around the cell perimeter further suggested that the mycobacterial sidewall is a site of cell wall assembly. While polar peptidoglycan synthesis was associated with cell elongation, sidewall synthesis responded to cell wall damage. Peptidoglycan editing along the sidewall may support cell wall robustness in pole-growing mycobacteria.
UR - https://elifesciences.org/articles/37243
M3 - Article
VL - 7
SP - e37243
JO - eLife
JF - eLife
SN - 2050-084X
ER -