Dataset Information

Results from a Multi-Laboratory Ocean Metaproteomic Intercomparison: Effects of LC-MS Acquisition and Data Analysis Procedures

ABSTRACT: Metaproteomics is an increasingly popular methodology that provides information regarding the metabolic functions of specific microbial taxa and can be used to assess environmental stressors and change and has potential for contributing to ocean ecology and biogeochemical studies. To enable future large-scale studies, a multi-laboratory intercomparison was conducted to assess comparability and reproducibility of taxonomic and functional results and their sensitivity to methodological variables. This ocean metaproteomic intercomparison consisted of two major activities: a laboratory component, where independent labs processed identical ocean samples simultaneously collected from the North Atlantic Ocean , and a subsequent informatic component.

INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Fusion, Q Exactive HF, Q Exactive

ORGANISM(S): Bacteria Environmental Samples <bacteria

DISEASE(S): Disease Free

SUBMITTER: Matthew McIlvin

LAB HEAD: Mak Saito

PROVIDER: PXD043218 | Pride | 2024-05-30

REPOSITORIES: Pride

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Source:

			Action	DRS
	Intercal_ORFs_prodigal_metagenemark.fasta	Fasta
	Intercal_assembly_annotations.csv	Csv
	Lab_127_1.mzid.gz	Mzid
	Lab_127_1.mzid_Lab_127_1.MGF	Mzid
	Lab_127_1.raw	Raw

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Project description:Metagenome-assembled genomes (MAGs) have revealed the existence of novel bacterial and archaeal groups and provided insight into their genetic potential. However, metagenomics and even metatranscriptomics cannot resolve how the genetic potential translates into metabolic functions and physiological activity. Here, we present a novel approach for the quantitative and organism-specific assessment of the carbon flux through microbial communities with stable isotope probing-metaproteomics and integration of temporal dynamics in 13C incorporation by Stable Isotope Cluster Analysis (SIsCA). We used groundwater microcosms labeled with 13CO2 and D2O as model systems and stimulated them with reduced sulfur compounds to determine the ecosystem role of chemolithoautotrophic primary production. Raman microspectroscopy detected rapid deuterium incorporation in microbial cells from 12 days onwards, indicating activity of the groundwater organisms. SIsCA revealed that groundwater microorganisms fell into five distinct carbon assimilation strategies. Only one of these strategies, comprising less than 3.5% of the community, consisted of obligate autotrophs (Thiobacillus), with a 13C incorporation of approximately 95%. Instead, mixotrophic growth was the most successful strategy, and was represented by 12 of the 15 MAGs expressing pathways for autotrophic CO2 fixation, including Hydrogenophaga, Polaromonas and Dechloromonas, with varying 13C incorporation between 5% and 90%. Within 21 days, 43% of carbon in the community was replaced by 13C, increasing to 80% after 70 days. Of the 31 most abundant MAGs, 16 expressed pathways for sulfur oxidation, including strict heterotrophs. We concluded that chemolithoautotrophy drives the recycling of organic carbon and serves as a fill-up function in the groundwater. Mixotrophs preferred the uptake of organic carbon over the fixation of CO2, and heterotrophs oxidize inorganic compounds to preserve organic carbon. Our study showcases how next-generation physiology approach like SIsCA can move beyond metagenomics studies by providing information about expression of metabolic pathways and elucidating the role of MAGs in ecosystem functioning.

Dataset Information

Results from a Multi-Laboratory Ocean Metaproteomic Intercomparison: Effects of LC-MS Acquisition and Data Analysis Procedures

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