TY - JOUR
T1 - Temporal Regulation of the Bacillus subtilis Acetylome and Evidence for a Role of MreB Acetylation in Cell Wall Growth
AU - Carabetta, Valerie J.
AU - Greco, Todd M.
AU - Tanner, Andrew W.
AU - Cristea, Ileana M.
AU - Dubnau, David
N1 - Funding Information:
We thank M. Neiditch for help with the structural modeling and discussions. A.W.T. was supported in part by the New Jersey Health Foundation. V.J.C., T.M.G., I.M.C., and D.D. designed the experimental strategy and experiments. V.J.C. performed the experiments, T.M.G. performed bioinformatic analyses, and A.W.T. performed statistical analysis. V.J.C., T.M.G., A.W.T., I.M.C., and D.D. wrote the manuscript. The authors declare that they have no conflicts of interest.
Funding Information:
This work, including the efforts of Todd M. Greco, was funded by NJCCR. This work, including the efforts of Ileana M. Cristea, was funded by HHS | National Institutes of Health (NIH) (R01GM114141, R21AI102187, and R01HL127640). This work, including the efforts of David Dubnau, was funded by HHS | National Institutes of Health (NIH) (GM057720 and GM043756).
Publisher Copyright:
Copyright © 2016 Carabetta et al.
PY - 2016/5/1
Y1 - 2016/5/1
N2 - N-Lysine acetylation has been recognized as a ubiquitous regulatory posttranslational modification that influences a variety of important biological processes in eukaryotic cells. Recently, it has been realized that acetylation is also prevalent in bacteria. Bacteria contain hundreds of acetylated proteins, with functions affecting diverse cellular pathways. Still, little is known about the regulation or biological relevance of nearly all of these modifications. Here we characterize the cellular growth-associated regulation of the Bacillus subtilis acetylome. Using acetylation enrichment and quantitative mass spectrometry, we investigate the logarithmic and stationary growth phases, identifying over 2,300 unique acetylation sites on proteins that function in essential cellular pathways. We determine an acetylation motif, EK(ac)(D/Y/E), which resembles the eukaryotic mitochondrial acetylation signature, and a distinct stationary-phase-enriched motif. By comparing the changes in acetylation with protein abundances, we discover a subset of critical acetylation events that are temporally regulated during cell growth. We functionally characterize the stationary-phase-enriched acetylation on the essential shape-determining protein MreB. Using bioinformatics, mutational analysis, and fluorescence microscopy, we define a potential role for the temporal acetylation of MreB in restricting cell wall growth and cell diameter. IMPORTANCE The past decade highlighted N-lysine acetylation as a prevalent posttranslational modification in bacteria. However, knowledge regarding the physiological importance and temporal regulation of acetylation has remained limited. To uncover potential regulatory roles for acetylation, we analyzed how acetylation patterns and abundances change between growth phases in B. subtilis. To demonstrate that the identification of cell growth-dependent modifications can point to critical regulatory acetylation events, we further characterized MreB, the cell shape-determining protein. Our findings led us to propose a role for MreB acetylation in controlling cell width by restricting cell wall growth.
AB - N-Lysine acetylation has been recognized as a ubiquitous regulatory posttranslational modification that influences a variety of important biological processes in eukaryotic cells. Recently, it has been realized that acetylation is also prevalent in bacteria. Bacteria contain hundreds of acetylated proteins, with functions affecting diverse cellular pathways. Still, little is known about the regulation or biological relevance of nearly all of these modifications. Here we characterize the cellular growth-associated regulation of the Bacillus subtilis acetylome. Using acetylation enrichment and quantitative mass spectrometry, we investigate the logarithmic and stationary growth phases, identifying over 2,300 unique acetylation sites on proteins that function in essential cellular pathways. We determine an acetylation motif, EK(ac)(D/Y/E), which resembles the eukaryotic mitochondrial acetylation signature, and a distinct stationary-phase-enriched motif. By comparing the changes in acetylation with protein abundances, we discover a subset of critical acetylation events that are temporally regulated during cell growth. We functionally characterize the stationary-phase-enriched acetylation on the essential shape-determining protein MreB. Using bioinformatics, mutational analysis, and fluorescence microscopy, we define a potential role for the temporal acetylation of MreB in restricting cell wall growth and cell diameter. IMPORTANCE The past decade highlighted N-lysine acetylation as a prevalent posttranslational modification in bacteria. However, knowledge regarding the physiological importance and temporal regulation of acetylation has remained limited. To uncover potential regulatory roles for acetylation, we analyzed how acetylation patterns and abundances change between growth phases in B. subtilis. To demonstrate that the identification of cell growth-dependent modifications can point to critical regulatory acetylation events, we further characterized MreB, the cell shape-determining protein. Our findings led us to propose a role for MreB acetylation in controlling cell width by restricting cell wall growth.
KW - Acetylome
KW - Cell wall synthesis
KW - Label-free quantification
KW - Lysine acetylation
KW - Mass spectrometry
KW - MreB
KW - Peptidoglycan
KW - Posttranslational modification
KW - Proteomics
UR - http://www.scopus.com/inward/record.url?scp=85006818296&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85006818296&partnerID=8YFLogxK
U2 - 10.1128/mSystems.00005-16
DO - 10.1128/mSystems.00005-16
M3 - Article
C2 - 27376153
AN - SCOPUS:85006818296
SN - 2379-5077
VL - 1
JO - mSystems
JF - mSystems
IS - 3
M1 - e00005-16
ER -