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Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1

Magnetotactic bacteria (MTB) are a phylogenetically diverse group of bacteria remarkable for their ability to biomineralize magnetite (Fe(3)O(4)) or greigite (Fe(3)S(4)) in organelles called magnetosomes. The majority of genes required for magnetosome formation are encoded by a magnetosome gene isla...

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Autores principales: McCausland, Hayley C., Wetmore, Kelly M., Arkin, Adam P., Komeili, Arash
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8788322/
https://www.ncbi.nlm.nih.gov/pubmed/35076272
http://dx.doi.org/10.1128/msystems.01037-21
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author McCausland, Hayley C.
Wetmore, Kelly M.
Arkin, Adam P.
Komeili, Arash
author_facet McCausland, Hayley C.
Wetmore, Kelly M.
Arkin, Adam P.
Komeili, Arash
author_sort McCausland, Hayley C.
collection PubMed
description Magnetotactic bacteria (MTB) are a phylogenetically diverse group of bacteria remarkable for their ability to biomineralize magnetite (Fe(3)O(4)) or greigite (Fe(3)S(4)) in organelles called magnetosomes. The majority of genes required for magnetosome formation are encoded by a magnetosome gene island (MAI). Most previous genetic studies of MTB have focused on the MAI, using screens to identify key MAI genes or targeted genetics to isolate specific genes and their function in one specific growth condition. This is the first study that has taken an unbiased approach to look at many different growth conditions to reveal key genes both inside and outside the MAI. Here, we conducted random barcoded transposon mutagenesis (RB-TnSeq) in Magnetospirillum magneticum AMB-1. We generated a library of 184,710 unique strains in a wild-type background, generating ∼34 mutant strains for each gene. RB-TnSeq also allowed us to determine the essential gene set of AMB-1 under standard laboratory growth conditions. To pinpoint novel genes that are important for magnetosome formation, we subjected the library to magnetic selection screens under varied growth conditions. We compared biomineralization under standard growth conditions to biomineralization under high-iron and anaerobic conditions, respectively. Strains with transposon insertions in the MAI gene mamT had an exacerbated biomineralization defect under both high-iron and anaerobic conditions compared to standard conditions, adding to our knowledge of the role of MamT in magnetosome formation. Mutants in an ex-MAI gene, amb4151, are more magnetic than wild-type cells under anaerobic conditions. All three of these phenotypes were validated by creating a markerless deletion strain of the gene and evaluating with TEM imaging. Overall, our results indicate that growth conditions affect which genes are required for biomineralization and that some MAI genes may have more nuanced functions than was previously understood. IMPORTANCE Magnetotactic bacteria (MTB) are a group of bacteria that can form nano-sized crystals of magnetic minerals. MTB are likely an important part of their ecosystems, because they can account for up to a third of the microbial biomass in an aquatic habitat and consume large amounts of iron, potentially impacting the iron cycle. The ecology of MTB is relatively understudied; however, the cell biology and genetics of MTB have been studied for decades. Here, we leverage genetic studies of MTB to inform environmental studies. We expand the genetic toolset for studying MTB in the lab and identify novel genes, or functions of genes, that have an impact on biomineralization.
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spelling pubmed-87883222022-02-07 Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1 McCausland, Hayley C. Wetmore, Kelly M. Arkin, Adam P. Komeili, Arash mSystems Research Article Magnetotactic bacteria (MTB) are a phylogenetically diverse group of bacteria remarkable for their ability to biomineralize magnetite (Fe(3)O(4)) or greigite (Fe(3)S(4)) in organelles called magnetosomes. The majority of genes required for magnetosome formation are encoded by a magnetosome gene island (MAI). Most previous genetic studies of MTB have focused on the MAI, using screens to identify key MAI genes or targeted genetics to isolate specific genes and their function in one specific growth condition. This is the first study that has taken an unbiased approach to look at many different growth conditions to reveal key genes both inside and outside the MAI. Here, we conducted random barcoded transposon mutagenesis (RB-TnSeq) in Magnetospirillum magneticum AMB-1. We generated a library of 184,710 unique strains in a wild-type background, generating ∼34 mutant strains for each gene. RB-TnSeq also allowed us to determine the essential gene set of AMB-1 under standard laboratory growth conditions. To pinpoint novel genes that are important for magnetosome formation, we subjected the library to magnetic selection screens under varied growth conditions. We compared biomineralization under standard growth conditions to biomineralization under high-iron and anaerobic conditions, respectively. Strains with transposon insertions in the MAI gene mamT had an exacerbated biomineralization defect under both high-iron and anaerobic conditions compared to standard conditions, adding to our knowledge of the role of MamT in magnetosome formation. Mutants in an ex-MAI gene, amb4151, are more magnetic than wild-type cells under anaerobic conditions. All three of these phenotypes were validated by creating a markerless deletion strain of the gene and evaluating with TEM imaging. Overall, our results indicate that growth conditions affect which genes are required for biomineralization and that some MAI genes may have more nuanced functions than was previously understood. IMPORTANCE Magnetotactic bacteria (MTB) are a group of bacteria that can form nano-sized crystals of magnetic minerals. MTB are likely an important part of their ecosystems, because they can account for up to a third of the microbial biomass in an aquatic habitat and consume large amounts of iron, potentially impacting the iron cycle. The ecology of MTB is relatively understudied; however, the cell biology and genetics of MTB have been studied for decades. Here, we leverage genetic studies of MTB to inform environmental studies. We expand the genetic toolset for studying MTB in the lab and identify novel genes, or functions of genes, that have an impact on biomineralization. American Society for Microbiology 2022-01-25 /pmc/articles/PMC8788322/ /pubmed/35076272 http://dx.doi.org/10.1128/msystems.01037-21 Text en Copyright © 2022 McCausland et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
McCausland, Hayley C.
Wetmore, Kelly M.
Arkin, Adam P.
Komeili, Arash
Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1
title Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1
title_full Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1
title_fullStr Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1
title_full_unstemmed Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1
title_short Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1
title_sort global analysis of biomineralization genes in magnetospirillum magneticum amb-1
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8788322/
https://www.ncbi.nlm.nih.gov/pubmed/35076272
http://dx.doi.org/10.1128/msystems.01037-21
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