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Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins
Bacterial signaling networks control a wide variety of cellular processes including growth, metabolism, and pathogenesis. Bis-(3′–5′)-cyclic dimeric guanosine monophosphate (cdiGMP) is a secondary signaling nucleotide that controls cellulose synthesis, biofilm formation, motility and virulence in a...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3538687/ https://www.ncbi.nlm.nih.gov/pubmed/23308275 http://dx.doi.org/10.1371/journal.pone.0053689 |
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author | Lieberman, Ori J. DeStefano, Jeffery J. Lee, Vincent T. |
author_facet | Lieberman, Ori J. DeStefano, Jeffery J. Lee, Vincent T. |
author_sort | Lieberman, Ori J. |
collection | PubMed |
description | Bacterial signaling networks control a wide variety of cellular processes including growth, metabolism, and pathogenesis. Bis-(3′–5′)-cyclic dimeric guanosine monophosphate (cdiGMP) is a secondary signaling nucleotide that controls cellulose synthesis, biofilm formation, motility and virulence in a wide range of Gram-negative bacterial species. CdiGMP is a dynamic molecule that forms different tertiary structures in vitro, including a trans-monomer, cis-monomer, cis-dimer and G-octaplex (G8). Although the monomer and dimer have been shown to be physiologically relevant in modulating protein activity and transcription, the biological effects of the cdiGMP G8 has not yet been described. Here, we have developed a TLC-based assay to detect radiolabeled cdiGMP G8 formation. Utilizing the radiolabeled cdiGMP G8, we have also shown a novel inhibitory interaction between the cdiGMP G8 and HIV-1 reverse transcriptase and that the cdiGMP G8 does not interact with proteins from Pseudomonas aeruginosa known to bind monomeric and dimeric cdiGMP. These results suggest that the radiolabeled cdiGMP G8 can be used to measure interactions between the cdiGMP G8 and cellular proteins, providing an avenue through which the biological significance of this molecule could be investigated. |
format | Online Article Text |
id | pubmed-3538687 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35386872013-01-10 Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins Lieberman, Ori J. DeStefano, Jeffery J. Lee, Vincent T. PLoS One Research Article Bacterial signaling networks control a wide variety of cellular processes including growth, metabolism, and pathogenesis. Bis-(3′–5′)-cyclic dimeric guanosine monophosphate (cdiGMP) is a secondary signaling nucleotide that controls cellulose synthesis, biofilm formation, motility and virulence in a wide range of Gram-negative bacterial species. CdiGMP is a dynamic molecule that forms different tertiary structures in vitro, including a trans-monomer, cis-monomer, cis-dimer and G-octaplex (G8). Although the monomer and dimer have been shown to be physiologically relevant in modulating protein activity and transcription, the biological effects of the cdiGMP G8 has not yet been described. Here, we have developed a TLC-based assay to detect radiolabeled cdiGMP G8 formation. Utilizing the radiolabeled cdiGMP G8, we have also shown a novel inhibitory interaction between the cdiGMP G8 and HIV-1 reverse transcriptase and that the cdiGMP G8 does not interact with proteins from Pseudomonas aeruginosa known to bind monomeric and dimeric cdiGMP. These results suggest that the radiolabeled cdiGMP G8 can be used to measure interactions between the cdiGMP G8 and cellular proteins, providing an avenue through which the biological significance of this molecule could be investigated. Public Library of Science 2013-01-07 /pmc/articles/PMC3538687/ /pubmed/23308275 http://dx.doi.org/10.1371/journal.pone.0053689 Text en © 2013 Lieberman et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Lieberman, Ori J. DeStefano, Jeffery J. Lee, Vincent T. Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins |
title | Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins |
title_full | Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins |
title_fullStr | Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins |
title_full_unstemmed | Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins |
title_short | Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins |
title_sort | detection of cyclic diguanylate g-octaplex assembly and interaction with proteins |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3538687/ https://www.ncbi.nlm.nih.gov/pubmed/23308275 http://dx.doi.org/10.1371/journal.pone.0053689 |
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