TY - JOUR
T1 - Binding of 2′,3′-Cyclic Nucleotide Monophosphates to Bacterial Ribosomes Inhibits Translation
AU - Chauhan, Shikha S.
AU - Marotta, Nick J.
AU - Karls, Anna C.
AU - Weinert, Emily E.
N1 - Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/11/23
Y1 - 2022/11/23
N2 - The intracellular small molecules 2′,3′-cyclic nucleotide monophosphates (2′,3′-cNMPs) have recently been rediscovered within both prokaryotes and eukaryotes. Studies in bacteria have demonstrated that 2′,3′-cNMP levels affect bacterial phenotypes, such as biofilm formation, motility, and growth, and modulate expression of numerous genes, suggesting that 2′,3′-cNMP levels are monitored by cells. In this study, 2′,3′-cNMP-linked affinity chromatography resins were used to identify Escherichia coli proteins that bind 2′,3′-cNMPs, with the top hits including all of the ribosomal proteins, and to confirm direct binding of purified ribosomes. Using in vitro translation assays, we have demonstrated that 2′,3′-cNMPs inhibit translation at concentrations found in amino acid-starved cells. In addition, a genetically encoded tool to increase cellular 2′,3′-cNMP levels was developed and was demonstrated to decrease E. coli growth rates. Taken together, this work suggests a mechanism for 2′,3-cNMP levels to modulate bacterial phenotypes by rapidly affecting translation.
AB - The intracellular small molecules 2′,3′-cyclic nucleotide monophosphates (2′,3′-cNMPs) have recently been rediscovered within both prokaryotes and eukaryotes. Studies in bacteria have demonstrated that 2′,3′-cNMP levels affect bacterial phenotypes, such as biofilm formation, motility, and growth, and modulate expression of numerous genes, suggesting that 2′,3′-cNMP levels are monitored by cells. In this study, 2′,3′-cNMP-linked affinity chromatography resins were used to identify Escherichia coli proteins that bind 2′,3′-cNMPs, with the top hits including all of the ribosomal proteins, and to confirm direct binding of purified ribosomes. Using in vitro translation assays, we have demonstrated that 2′,3′-cNMPs inhibit translation at concentrations found in amino acid-starved cells. In addition, a genetically encoded tool to increase cellular 2′,3′-cNMP levels was developed and was demonstrated to decrease E. coli growth rates. Taken together, this work suggests a mechanism for 2′,3-cNMP levels to modulate bacterial phenotypes by rapidly affecting translation.
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U2 - 10.1021/acscentsci.2c00681
DO - 10.1021/acscentsci.2c00681
M3 - Article
C2 - 36439312
AN - SCOPUS:85139506902
SN - 2374-7943
VL - 8
SP - 1518
EP - 1526
JO - ACS Central Science
JF - ACS Central Science
IS - 11
ER -