Protein acetylation dynamics in response to carbon overflow in Escherichia coli.
In Escherichia coli, acetylation of proteins at lysines depends largely on a non-enzymatic acetyl phosphate-dependent mechanism. To assess the functional significance of this post-translational modification, we first grew wild-type cells in buffered tryptone broth with glucose and monitored acetylation over time by immunochemistry. Most acetylation occurred in stationary phase and paralleled glucose consumption and acetate excretion, which began upon entry into stationary phase. Transcription of rprA, a stationary phase regulator, exhibited similar behavior. To identify sites and substrates with significant acetylation changes, we used label-free, quantitative proteomics to monitor changes in protein acetylation. During growth, both the number of identified sites and the extent of acetylation increased with considerable variation among lysines from the same protein. As glucose-regulated lysine acetylation was predominant in central metabolic pathways and overlapped with acetyl phosphate-regulated acetylation sites, we deleted the major carbon regulator CRP and observed a dramatic loss of acetylation that could be restored by deleting the enzyme that degrades acetyl phosphate. We propose that acetyl phosphate-dependent acetylation is a response to carbon flux that could regulate central metabolism.
Buck Institute for Research on Aging, Novato, CA, 94945, USA.
Birgit
B
Schilling
Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL, 60153, USA.
David
D
Christensen
Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL, 60153, USA.
Robert
R
Davis
Buck Institute for Research on Aging, Novato, CA, 94945, USA.
Alexandria K
AK
Sahu
Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL, 60153, USA.
Linda I
LI
Hu
Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL, 60153, USA.
Arti
A
Walker-Peddakotla
Buck Institute for Research on Aging, Novato, CA, 94945, USA.
Dylan J
DJ
Sorensen
Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL, 60153, USA.
Bozena
B
Zemaitaitis
Buck Institute for Research on Aging, Novato, CA, 94945, USA.|Department of Pharmaceutical Chemistry, University of California, San Francisco, CA, 94143, USA.
Bradford W
BW
Gibson
Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL, 60153, USA.
Alan J
AJ
Wolfe
D000085:Acetates / Q000378:metabolism; D000107:Acetylation; D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D002244:Carbon / Q000378:metabolism*; D057486:Carbon Cycle / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D011499:Protein Processing, Post-Translational; D040901:Proteomics
D000085:Acetates; D029968:Escherichia coli Proteins; D002244:Carbon; D000123:Acetyltransferases; D005947:Glucose; D008239:Lysine