Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase
Mammalian cyclic GMP-AMP synthase (cGAS) and its homologue dinucleotide cyclase in Vibrio cholerae (VcDncV) produce cyclic dinucleotides (CDNs) that participate in the defense against viral infection. Recently, scores of new cGAS/DncV-like nucleotidyltransferases (CD-NTases) were discovered, which p...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Oxford University Press
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8096243/ https://www.ncbi.nlm.nih.gov/pubmed/33836064 http://dx.doi.org/10.1093/nar/gkab165 |
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author | Ko, Tzu-Ping Wang, Yu-Chuan Tsai, Chia-Ling Yang, Chia-Shin Hou, Mei-Hui Chen, Yeh |
author_facet | Ko, Tzu-Ping Wang, Yu-Chuan Tsai, Chia-Ling Yang, Chia-Shin Hou, Mei-Hui Chen, Yeh |
author_sort | Ko, Tzu-Ping |
collection | PubMed |
description | Mammalian cyclic GMP-AMP synthase (cGAS) and its homologue dinucleotide cyclase in Vibrio cholerae (VcDncV) produce cyclic dinucleotides (CDNs) that participate in the defense against viral infection. Recently, scores of new cGAS/DncV-like nucleotidyltransferases (CD-NTases) were discovered, which produce various CDNs and cyclic trinucleotides (CTNs) as second messengers. Here, we present the crystal structures of EcCdnD, a CD-NTase from Enterobacter cloacae that produces cyclic AMP-AMP-GMP, in its apo-form and in complex with ATP, ADP and AMPcPP, an ATP analogue. Despite the similar overall architecture, the protein shows significant structural variations from other CD-NTases. Adjacent to the donor substrate, another nucleotide is bound to the acceptor binding site by a non-productive mode. Isothermal titration calorimetry results also suggest the presence of two ATP binding sites. GTP alone does not bind to EcCdnD, which however binds to pppApG, a possible intermediate. The enzyme is active on ATP or a mixture of ATP and GTP, and the best metal cofactor is Mg(2+). The conserved residues Asp69 and Asp71 are essential for catalysis, as indicated by the loss of activity in the mutants. Based on structural analysis and comparison with VcDncV and RNA polymerase, a tentative catalytic pathway for the CTN-producing EcCdnD is proposed. |
format | Online Article Text |
id | pubmed-8096243 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-80962432021-05-10 Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase Ko, Tzu-Ping Wang, Yu-Chuan Tsai, Chia-Ling Yang, Chia-Shin Hou, Mei-Hui Chen, Yeh Nucleic Acids Res Structural Biology Mammalian cyclic GMP-AMP synthase (cGAS) and its homologue dinucleotide cyclase in Vibrio cholerae (VcDncV) produce cyclic dinucleotides (CDNs) that participate in the defense against viral infection. Recently, scores of new cGAS/DncV-like nucleotidyltransferases (CD-NTases) were discovered, which produce various CDNs and cyclic trinucleotides (CTNs) as second messengers. Here, we present the crystal structures of EcCdnD, a CD-NTase from Enterobacter cloacae that produces cyclic AMP-AMP-GMP, in its apo-form and in complex with ATP, ADP and AMPcPP, an ATP analogue. Despite the similar overall architecture, the protein shows significant structural variations from other CD-NTases. Adjacent to the donor substrate, another nucleotide is bound to the acceptor binding site by a non-productive mode. Isothermal titration calorimetry results also suggest the presence of two ATP binding sites. GTP alone does not bind to EcCdnD, which however binds to pppApG, a possible intermediate. The enzyme is active on ATP or a mixture of ATP and GTP, and the best metal cofactor is Mg(2+). The conserved residues Asp69 and Asp71 are essential for catalysis, as indicated by the loss of activity in the mutants. Based on structural analysis and comparison with VcDncV and RNA polymerase, a tentative catalytic pathway for the CTN-producing EcCdnD is proposed. Oxford University Press 2021-04-09 /pmc/articles/PMC8096243/ /pubmed/33836064 http://dx.doi.org/10.1093/nar/gkab165 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/ (https://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 | Structural Biology Ko, Tzu-Ping Wang, Yu-Chuan Tsai, Chia-Ling Yang, Chia-Shin Hou, Mei-Hui Chen, Yeh Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase |
title | Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase |
title_full | Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase |
title_fullStr | Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase |
title_full_unstemmed | Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase |
title_short | Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase |
title_sort | crystal structure and functional implication of a bacterial cyclic amp–amp–gmp synthetase |
topic | Structural Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8096243/ https://www.ncbi.nlm.nih.gov/pubmed/33836064 http://dx.doi.org/10.1093/nar/gkab165 |
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