TY - JOUR
T1 - Saltinduced stress stimulates a lipoteichoic acidspecific three-component glycosylation system in Staphylococcus aureus
AU - Kho, Kelvin
AU - Meredith, Timothy C.
N1 - Funding Information:
We thank Gloria Komazin for help with cloning and strain construction (Pennsylvania State University), Tapas Mal, Carlos Pacheco, and Debashish Sahu (Penn State NMR Facility, University Park, PA) for expert technical assistance with NMR, James R. Miller for expert technical assistance with ESI-MS (Penn State Proteomics and Mass Spectrometry Core Facility, University Park, PA), and Suzanne Walker for strains TM226 and SHM084
Publisher Copyright:
© 2018 American Society for Microbiology.
PY - 2018/6/1
Y1 - 2018/6/1
N2 - Lipoteichoic acid (LTA) in Staphylococcus aureus is a poly-glycerophosphate polymer anchored to the outer surface of the cell membrane. LTA has numerous roles in cell envelope physiology, including regulating cell autolysis, coordinating cell division, and adapting to environmental growth conditions. LTA is often further modified with substituents, including D-alanine and glycosyl groups, to alter cellular function. While the genetic determinants of D-alanylation have been largely defined, the route of LTA glycosylation and its role in cell envelope physiology have remained unknown, in part due to the low levels of basal LTA glycosylation in S. aureus. We demonstrate here that S. aureus utilizes a membrane-associated three-component glycosylation system composed of an undecaprenol (Und) N-acetylglucosamine (GlcNAc) charging enzyme (CsbB; SAOUHSC_00713), a putative flippase to transport loaded substrate to the outside surface of the cell (GtcA; SAOUHSC_02722), and finally an LTA-specific glycosyltransferase that adds α-GlcNAc moieties to LTA (YfhO; SAOUHSC_01213). We demonstrate that this system is specific for LTA with no cross recognition of the structurally similar polyribitol phosphate containing wall teichoic acids. We show that while wild-type S. aureus LTA has only a trace of GlcNAcylated LTA under normal growth conditions, amounts are raised upon either overexpressing CsbB, reducing endogenous D-alanylation activity, expressing the cell envelope stress responsive alternative sigma factor SigB, or by exposure to environmental stress-inducing culture conditions, including growth media containing high levels of sodium chloride.
AB - Lipoteichoic acid (LTA) in Staphylococcus aureus is a poly-glycerophosphate polymer anchored to the outer surface of the cell membrane. LTA has numerous roles in cell envelope physiology, including regulating cell autolysis, coordinating cell division, and adapting to environmental growth conditions. LTA is often further modified with substituents, including D-alanine and glycosyl groups, to alter cellular function. While the genetic determinants of D-alanylation have been largely defined, the route of LTA glycosylation and its role in cell envelope physiology have remained unknown, in part due to the low levels of basal LTA glycosylation in S. aureus. We demonstrate here that S. aureus utilizes a membrane-associated three-component glycosylation system composed of an undecaprenol (Und) N-acetylglucosamine (GlcNAc) charging enzyme (CsbB; SAOUHSC_00713), a putative flippase to transport loaded substrate to the outside surface of the cell (GtcA; SAOUHSC_02722), and finally an LTA-specific glycosyltransferase that adds α-GlcNAc moieties to LTA (YfhO; SAOUHSC_01213). We demonstrate that this system is specific for LTA with no cross recognition of the structurally similar polyribitol phosphate containing wall teichoic acids. We show that while wild-type S. aureus LTA has only a trace of GlcNAcylated LTA under normal growth conditions, amounts are raised upon either overexpressing CsbB, reducing endogenous D-alanylation activity, expressing the cell envelope stress responsive alternative sigma factor SigB, or by exposure to environmental stress-inducing culture conditions, including growth media containing high levels of sodium chloride.
UR - http://www.scopus.com/inward/record.url?scp=85047467770&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85047467770&partnerID=8YFLogxK
U2 - 10.1128/JB.00017-18
DO - 10.1128/JB.00017-18
M3 - Article
C2 - 29632092
AN - SCOPUS:85047467770
SN - 0021-9193
VL - 200
JO - Journal of bacteriology
JF - Journal of bacteriology
IS - 12
M1 - e00017.18
ER -