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Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology

Bacterial micro-compartments (BMC) are complex macromolecular assemblies that participate in varied metabolic processes in about 20% of bacterial species. Most of these organisms carry BMC genetic information organized in operons that often include several paralog genes coding for components of the...

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Autores principales: Barthe, Lucie, Soldan, Vanessa, Garcia-Alles, Luis F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10681173/
https://www.ncbi.nlm.nih.gov/pubmed/38011088
http://dx.doi.org/10.1371/journal.pone.0294760
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author Barthe, Lucie
Soldan, Vanessa
Garcia-Alles, Luis F.
author_facet Barthe, Lucie
Soldan, Vanessa
Garcia-Alles, Luis F.
author_sort Barthe, Lucie
collection PubMed
description Bacterial micro-compartments (BMC) are complex macromolecular assemblies that participate in varied metabolic processes in about 20% of bacterial species. Most of these organisms carry BMC genetic information organized in operons that often include several paralog genes coding for components of the compartment shell. BMC shell constituents can be classified depending on their oligomerization state as hexamers (BMC-H), pentamers (BMC-P) or trimers (BMC-T). Formation of hetero-oligomers combining different protein homologs is theoretically feasible, something that could ultimately modify BMC shell rigidity or permeability, for instance. Despite that, it remains largely unknown whether hetero-oligomerization is a widespread phenomenon. Here, we demonstrated that the tripartite GFP (tGFP) reporter technology is an appropriate tool that might be exploited for such purposes. Thus, after optimizing parameters such as the size of linkers connecting investigated proteins to GFP10 or GFP11 peptides, the type and strength of promoters, or the impact of placing coding cassettes in the same or different plasmids, homo-oligomerization processes could be successfully monitored for any of the three BMC shell classes. Moreover, the screen perfectly reproduced published data on hetero-association between couples of CcmK homologues from Syn. sp. PCC6803, which were obtained following a different approach. This study paves the way for mid/high throughput screens to characterize the extent of hetero-oligomerization occurrence in BMC-possessing bacteria, and most especially in organisms endowed with several BMC types and carrying numerous shell paralogs. On the other hand, our study also unveiled technology limitations deriving from the low solubility of one of the components of this modified split-GFP approach, the GFP1-9.
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spelling pubmed-106811732023-11-27 Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology Barthe, Lucie Soldan, Vanessa Garcia-Alles, Luis F. PLoS One Research Article Bacterial micro-compartments (BMC) are complex macromolecular assemblies that participate in varied metabolic processes in about 20% of bacterial species. Most of these organisms carry BMC genetic information organized in operons that often include several paralog genes coding for components of the compartment shell. BMC shell constituents can be classified depending on their oligomerization state as hexamers (BMC-H), pentamers (BMC-P) or trimers (BMC-T). Formation of hetero-oligomers combining different protein homologs is theoretically feasible, something that could ultimately modify BMC shell rigidity or permeability, for instance. Despite that, it remains largely unknown whether hetero-oligomerization is a widespread phenomenon. Here, we demonstrated that the tripartite GFP (tGFP) reporter technology is an appropriate tool that might be exploited for such purposes. Thus, after optimizing parameters such as the size of linkers connecting investigated proteins to GFP10 or GFP11 peptides, the type and strength of promoters, or the impact of placing coding cassettes in the same or different plasmids, homo-oligomerization processes could be successfully monitored for any of the three BMC shell classes. Moreover, the screen perfectly reproduced published data on hetero-association between couples of CcmK homologues from Syn. sp. PCC6803, which were obtained following a different approach. This study paves the way for mid/high throughput screens to characterize the extent of hetero-oligomerization occurrence in BMC-possessing bacteria, and most especially in organisms endowed with several BMC types and carrying numerous shell paralogs. On the other hand, our study also unveiled technology limitations deriving from the low solubility of one of the components of this modified split-GFP approach, the GFP1-9. Public Library of Science 2023-11-27 /pmc/articles/PMC10681173/ /pubmed/38011088 http://dx.doi.org/10.1371/journal.pone.0294760 Text en © 2023 Barthe et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Barthe, Lucie
Soldan, Vanessa
Garcia-Alles, Luis F.
Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology
title Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology
title_full Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology
title_fullStr Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology
title_full_unstemmed Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology
title_short Assessment of oligomerization of bacterial micro-compartment shell components with the tripartite GFP reporter technology
title_sort assessment of oligomerization of bacterial micro-compartment shell components with the tripartite gfp reporter technology
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10681173/
https://www.ncbi.nlm.nih.gov/pubmed/38011088
http://dx.doi.org/10.1371/journal.pone.0294760
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