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 - 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
SN - 2058-5276
VL - 6
SP - 584
EP - 593
JO - Nature microbiology
JF - Nature microbiology
IS - 5
ER -