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Deacetylation of topoisomerase I is an important physiological function of E. coli CobB

Escherichia coli topoisomerase I (TopA), a regulator of global and local DNA supercoiling, is modified by N(ε)-Lysine acetylation. The NAD(+)-dependent protein deacetylase CobB can reverse both enzymatic and non-enzymatic lysine acetylation modification in E. coli. Here, we show that the absence of...

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Autores principales: Zhou, Qingxuan, Zhou, Yan Ning, Jin, Ding Jun, Tse-Dinh, Yuk-Ching
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5605244/
https://www.ncbi.nlm.nih.gov/pubmed/28398568
http://dx.doi.org/10.1093/nar/gkx250
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author Zhou, Qingxuan
Zhou, Yan Ning
Jin, Ding Jun
Tse-Dinh, Yuk-Ching
author_facet Zhou, Qingxuan
Zhou, Yan Ning
Jin, Ding Jun
Tse-Dinh, Yuk-Ching
author_sort Zhou, Qingxuan
collection PubMed
description Escherichia coli topoisomerase I (TopA), a regulator of global and local DNA supercoiling, is modified by N(ε)-Lysine acetylation. The NAD(+)-dependent protein deacetylase CobB can reverse both enzymatic and non-enzymatic lysine acetylation modification in E. coli. Here, we show that the absence of CobB in a ΔcobB mutant reduces intracellular TopA catalytic activity and increases negative DNA supercoiling. TopA expression level is elevated as topA transcription responds to the increased negative supercoiling. The slow growth phenotype of the ΔcobB mutant can be partially compensated by further increase of intracellular TopA level via overexpression of recombinant TopA. The relaxation activity of purified TopA is decreased by in vitro non-enzymatic acetyl phosphate mediated lysine acetylation, and the presence of purified CobB protects TopA from inactivation by such non-enzymatic acetylation. The specific activity of TopA expressed from His-tagged fusion construct in the chromosome is inversely proportional to the degree of in vivo lysine acetylation during growth transition and growth arrest. These findings demonstrate that E. coli TopA catalytic activity can be modulated by lysine acetylation–deacetylation, and prevention of TopA inactivation from excess lysine acetylation and consequent increase in negative DNA supercoiling is an important physiological function of the CobB protein deacetylase.
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spelling pubmed-56052442017-09-25 Deacetylation of topoisomerase I is an important physiological function of E. coli CobB Zhou, Qingxuan Zhou, Yan Ning Jin, Ding Jun Tse-Dinh, Yuk-Ching Nucleic Acids Res Nucleic Acid Enzymes Escherichia coli topoisomerase I (TopA), a regulator of global and local DNA supercoiling, is modified by N(ε)-Lysine acetylation. The NAD(+)-dependent protein deacetylase CobB can reverse both enzymatic and non-enzymatic lysine acetylation modification in E. coli. Here, we show that the absence of CobB in a ΔcobB mutant reduces intracellular TopA catalytic activity and increases negative DNA supercoiling. TopA expression level is elevated as topA transcription responds to the increased negative supercoiling. The slow growth phenotype of the ΔcobB mutant can be partially compensated by further increase of intracellular TopA level via overexpression of recombinant TopA. The relaxation activity of purified TopA is decreased by in vitro non-enzymatic acetyl phosphate mediated lysine acetylation, and the presence of purified CobB protects TopA from inactivation by such non-enzymatic acetylation. The specific activity of TopA expressed from His-tagged fusion construct in the chromosome is inversely proportional to the degree of in vivo lysine acetylation during growth transition and growth arrest. These findings demonstrate that E. coli TopA catalytic activity can be modulated by lysine acetylation–deacetylation, and prevention of TopA inactivation from excess lysine acetylation and consequent increase in negative DNA supercoiling is an important physiological function of the CobB protein deacetylase. Oxford University Press 2017-05-19 2017-04-08 /pmc/articles/PMC5605244/ /pubmed/28398568 http://dx.doi.org/10.1093/nar/gkx250 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Nucleic Acid Enzymes
Zhou, Qingxuan
Zhou, Yan Ning
Jin, Ding Jun
Tse-Dinh, Yuk-Ching
Deacetylation of topoisomerase I is an important physiological function of E. coli CobB
title Deacetylation of topoisomerase I is an important physiological function of E. coli CobB
title_full Deacetylation of topoisomerase I is an important physiological function of E. coli CobB
title_fullStr Deacetylation of topoisomerase I is an important physiological function of E. coli CobB
title_full_unstemmed Deacetylation of topoisomerase I is an important physiological function of E. coli CobB
title_short Deacetylation of topoisomerase I is an important physiological function of E. coli CobB
title_sort deacetylation of topoisomerase i is an important physiological function of e. coli cobb
topic Nucleic Acid Enzymes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5605244/
https://www.ncbi.nlm.nih.gov/pubmed/28398568
http://dx.doi.org/10.1093/nar/gkx250
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