TY - JOUR
T1 - A two-track model for the spatiotemporal coordination of bacterial septal cell wall synthesis revealed by single-molecule imaging of FtsW
AU - Yang, Xinxing
AU - McQuillen, Ryan
AU - Lyu, Zhixin
AU - Phillips-Mason, Polly
AU - De La Cruz, Ana
AU - McCausland, Joshua W.
AU - Liang, Hai
AU - DeMeester, Kristen E.
AU - Santiago, Cintia C.
AU - Grimes, Catherine L.
AU - de Boer, Piet
AU - Xiao, Jie
N1 - Funding Information:
We thank other laboratory members in the Xiao and de Boer laboratories for helpful discussions and technical assistance. We also thank G. Hauk for sharing plasmids and the CRISPR-Cas9/λ-red recombineering cloning method, D. S. Weiss for strain EC1908, plasmid pDSW406, anti-FtsN serum and helpful suggestions on FtsW immunoblotting, T. Bernhardt for strain HC532 and plasmid pHC808, C. Hale for plasmid pCH650, E. Goley for help on cell growth measurement and R. Tsien for the TagRFP-T construct. This work was supported by NIH U01CA221230 (to C.L.G.), Pew Biomedical Scholar (Pew Foundation to C.L.G.), NIH T32GM133395A (to K.E.D.), NIH GM57059 (to P.d.B.), NIH R01GM086447 and R35GM136436 (to J.X.), GM125656 (subcontract to J.X.), NSF EAGER award MCB-1019000 (to J.X.) and a Hamilton Smith Innovative Research Award (to J.X.).
Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer Nature Limited.
PY - 2021/5
Y1 - 2021/5
N2 - Synthesis of septal peptidoglycan (sPG) is crucial for bacterial cell division. FtsW, an indispensable component of the cell division machinery in all walled bacterial species, was recently identified in vitro as a peptidoglycan glycosyltransferase (PGTase). Despite its importance, the septal PGTase activity of FtsW has not been demonstrated in vivo. How its activity is spatiotemporally regulated in vivo has also remained elusive. Here, we confirmed FtsW as an essential septum-specific PGTase in vivo using an N-acetylmuramic acid analogue incorporation assay. Next, using single-molecule tracking coupled with genetic manipulations, we identified two populations of processively moving FtsW molecules: a fast-moving population correlated with the treadmilling dynamics of the essential cytoskeletal FtsZ protein and a slow-moving population dependent on active sPG synthesis. We further identified that FtsN, a potential sPG synthesis activator, plays an important role in promoting the slow-moving population. Our results suggest a two-track model, in which inactive sPG synthases follow the ‘Z-track’ to be distributed along the septum and FtsN promotes their release from the Z-track to become active in sPG synthesis on the slow ‘sPG-track’. This model provides a mechanistic framework for the spatiotemporal coordination of sPG synthesis in bacterial cell division.
AB - Synthesis of septal peptidoglycan (sPG) is crucial for bacterial cell division. FtsW, an indispensable component of the cell division machinery in all walled bacterial species, was recently identified in vitro as a peptidoglycan glycosyltransferase (PGTase). Despite its importance, the septal PGTase activity of FtsW has not been demonstrated in vivo. How its activity is spatiotemporally regulated in vivo has also remained elusive. Here, we confirmed FtsW as an essential septum-specific PGTase in vivo using an N-acetylmuramic acid analogue incorporation assay. Next, using single-molecule tracking coupled with genetic manipulations, we identified two populations of processively moving FtsW molecules: a fast-moving population correlated with the treadmilling dynamics of the essential cytoskeletal FtsZ protein and a slow-moving population dependent on active sPG synthesis. We further identified that FtsN, a potential sPG synthesis activator, plays an important role in promoting the slow-moving population. Our results suggest a two-track model, in which inactive sPG synthases follow the ‘Z-track’ to be distributed along the septum and FtsN promotes their release from the Z-track to become active in sPG synthesis on the slow ‘sPG-track’. This model provides a mechanistic framework for the spatiotemporal coordination of sPG synthesis in bacterial cell division.
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U2 - 10.1038/s41564-020-00853-0
DO - 10.1038/s41564-020-00853-0
M3 - Article
C2 - 33495624
AN - SCOPUS:85099760113
VL - 6
SP - 584
EP - 593
JO - Nature Microbiology
JF - Nature Microbiology
SN - 2058-5276
IS - 5
ER -