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Title: Metabolic Network Analysis and Metatranscriptomics Reveal Auxotrophies and Nutrient Sources of the Cosmopolitan Freshwater Microbial Lineage acI

Abstract

An explosion in the number of available genome sequences obtained through metagenomics and single-cell genomics has enabled a new view of the diversity of microbial life, yet we know surprisingly little about how microbes interact with each other or their environment. In fact, the majority of microbial species remain uncultivated, while our perception of their ecological niches is based on reconstruction of their metabolic potential. In this work, we demonstrate how the “seed set framework,” which computes the set of compounds that an organism must acquire from its environment (E. Borenstein, M. Kupiec, M. W. Feldman, and E. Ruppin, Proc Natl Acad Sci U S A 105:14482–14487, 2008, https://doi.org/10.1073/pnas.0806162105), enables computational analysis of metabolic reconstructions while providing new insights into a microbe’s metabolic capabilities, such as nutrient use and auxotrophies. We apply this framework to members of the ubiquitous freshwater actinobacterial lineage acI, confirming and extending previous experimental and genomic observations implying that acI bacteria are heterotrophs reliant on peptides and saccharides. We also present the first metatranscriptomic study of the acI lineage, revealing high expression of transport proteins and the light-harvesting protein actinorhodopsin. Putative transport proteins complement predictions of nutrients and essential metabolites while providing additional support of themore » hypothesis that members of the acI are photoheterotrophs.« less

Authors:
ORCiD logo [1]; ORCiD logo [2];  [1]; ORCiD logo [3];  [3]; ORCiD logo [4]; ORCiD logo [5];  [1]; ORCiD logo [6]
  1. Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology
  2. Uppsala Univ. (Sweden). Dept. of Ecology and Genetics
  3. Univ. of Wisconsin, Madison, WI (United States). Dept. of Civil and Environmental Engineering
  4. Uppsala Univ. (Sweden). Dept. of Ecology and Genetics. Science for Life Lab.
  5. USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
  6. Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology. Dept. of Civil and Environmental Engineering
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); USDA National Inst. of Food and Agriculture (NIFA); National Science Foundation (NSF); National Oceanic and Atmospheric Administration (NOAA) (United States)
OSTI Identifier:
1459392
Grant/Contract Number:  
AC02-05CH11231; 2016-67012-24709; WIS01789; MCB-0702395; NTL-LTER DEB-1440297; DEB-1344254; NA10OAR4170070
Resource Type:
Accepted Manuscript
Journal Name:
mSystems
Additional Journal Information:
Journal Volume: 2; Journal Issue: 4; Journal ID: ISSN 2379-5077
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; freshwater microbial ecology; metabolism; metagenomics; metatranscriptomics; physiology; systems biology

Citation Formats

Hamilton, Joshua J., Garcia, Sarahi L., Brown, Brittany S., Oyserman, Ben O., Moya-Flores, Francisco, Bertilsson, Stefan, Malmstrom, Rex R., Forest, Katrina T., and McMahon, Katherine D. Metabolic Network Analysis and Metatranscriptomics Reveal Auxotrophies and Nutrient Sources of the Cosmopolitan Freshwater Microbial Lineage acI. United States: N. p., 2017. Web. doi:10.1128/mSystems.00091-17.
Hamilton, Joshua J., Garcia, Sarahi L., Brown, Brittany S., Oyserman, Ben O., Moya-Flores, Francisco, Bertilsson, Stefan, Malmstrom, Rex R., Forest, Katrina T., & McMahon, Katherine D. Metabolic Network Analysis and Metatranscriptomics Reveal Auxotrophies and Nutrient Sources of the Cosmopolitan Freshwater Microbial Lineage acI. United States. https://doi.org/10.1128/mSystems.00091-17
Hamilton, Joshua J., Garcia, Sarahi L., Brown, Brittany S., Oyserman, Ben O., Moya-Flores, Francisco, Bertilsson, Stefan, Malmstrom, Rex R., Forest, Katrina T., and McMahon, Katherine D. Tue . "Metabolic Network Analysis and Metatranscriptomics Reveal Auxotrophies and Nutrient Sources of the Cosmopolitan Freshwater Microbial Lineage acI". United States. https://doi.org/10.1128/mSystems.00091-17. https://www.osti.gov/servlets/purl/1459392.
@article{osti_1459392,
title = {Metabolic Network Analysis and Metatranscriptomics Reveal Auxotrophies and Nutrient Sources of the Cosmopolitan Freshwater Microbial Lineage acI},
author = {Hamilton, Joshua J. and Garcia, Sarahi L. and Brown, Brittany S. and Oyserman, Ben O. and Moya-Flores, Francisco and Bertilsson, Stefan and Malmstrom, Rex R. and Forest, Katrina T. and McMahon, Katherine D.},
abstractNote = {An explosion in the number of available genome sequences obtained through metagenomics and single-cell genomics has enabled a new view of the diversity of microbial life, yet we know surprisingly little about how microbes interact with each other or their environment. In fact, the majority of microbial species remain uncultivated, while our perception of their ecological niches is based on reconstruction of their metabolic potential. In this work, we demonstrate how the “seed set framework,” which computes the set of compounds that an organism must acquire from its environment (E. Borenstein, M. Kupiec, M. W. Feldman, and E. Ruppin, Proc Natl Acad Sci U S A 105:14482–14487, 2008, https://doi.org/10.1073/pnas.0806162105), enables computational analysis of metabolic reconstructions while providing new insights into a microbe’s metabolic capabilities, such as nutrient use and auxotrophies. We apply this framework to members of the ubiquitous freshwater actinobacterial lineage acI, confirming and extending previous experimental and genomic observations implying that acI bacteria are heterotrophs reliant on peptides and saccharides. We also present the first metatranscriptomic study of the acI lineage, revealing high expression of transport proteins and the light-harvesting protein actinorhodopsin. Putative transport proteins complement predictions of nutrients and essential metabolites while providing additional support of the hypothesis that members of the acI are photoheterotrophs.},
doi = {10.1128/mSystems.00091-17},
journal = {mSystems},
number = 4,
volume = 2,
place = {United States},
year = {Tue Aug 29 00:00:00 EDT 2017},
month = {Tue Aug 29 00:00:00 EDT 2017}
}

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Works referenced in this record:

The Microbial Engines That Drive Earth's Biogeochemical Cycles
journal, May 2008


Toward a Predictive Understanding of Earth’s Microbiomes to Address 21st Century Challenges
journal, May 2016


Recovering complete and draft population genomes from metagenome datasets
journal, March 2016


Metabolic network analysis reveals microbial community interactions in anammox granules
journal, May 2017

  • Lawson, Christopher E.; Wu, Sha; Bhattacharjee, Ananda S.
  • Nature Communications, Vol. 8, Issue 1
  • DOI: 10.1038/ncomms15416

Thousands of microbial genomes shed light on interconnected biogeochemical processes in an aquifer system
journal, October 2016

  • Anantharaman, Karthik; Brown, Christopher T.; Hug, Laura A.
  • Nature Communications, Vol. 7, Issue 1
  • DOI: 10.1038/ncomms13219

What is flux balance analysis?
journal, March 2010

  • Orth, Jeffrey D.; Thiele, Ines; Palsson, Bernhard Ø
  • Nature Biotechnology, Vol. 28, Issue 3
  • DOI: 10.1038/nbt.1614

Large-scale reconstruction and phylogenetic analysis of metabolic environments
journal, September 2008

  • Borenstein, E.; Kupiec, M.; Feldman, M. W.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 38
  • DOI: 10.1073/pnas.0806162105

Nearly Identical 16S rRNA Sequences Recovered from Lakes in North America and Europe Indicate the Existence of Clades of Globally Distributed Freshwater Bacteria
journal, December 1998

  • Zwart, Gabriel; Hiorns, William D.; Methé, Barbara A.
  • Systematic and Applied Microbiology, Vol. 21, Issue 4
  • DOI: 10.1016/S0723-2020(98)80067-2

Comparative 16S rRNA Analysis of Lake Bacterioplankton Reveals Globally Distributed Phylogenetic Clusters Including an Abundant Group of Actinobacteria
journal, November 2000


Typical freshwater bacteria: an analysis of available 16S rRNA gene sequences from plankton of lakes and rivers
journal, January 2002

  • Zwart, G.; Crump, Bc; Kamst-van Agterveld, Mp
  • Aquatic Microbial Ecology, Vol. 28
  • DOI: 10.3354/ame028141

Microbial community dynamics in a humic lake: differential persistence of common freshwater phylotypes
journal, June 2006


Bacterioplankton Community Composition along a Salinity Gradient of Sixteen High-Mountain Lakes Located on the Tibetan Plateau, China
journal, August 2006

  • Wu, Q. L.; Zwart, G.; Schauer, M.
  • Applied and Environmental Microbiology, Vol. 72, Issue 8
  • DOI: 10.1128/AEM.00767-06

Phylogenetic Ecology of the Freshwater Actinobacteria acI Lineage
journal, September 2007

  • Newton, R. J.; Jones, S. E.; Helmus, M. R.
  • Applied and Environmental Microbiology, Vol. 73, Issue 22
  • DOI: 10.1128/AEM.00794-07

Clone library analysis reveals an unusual composition and strong habitat partitioning of pelagic bacterial communities in Lake Tanganyika
journal, January 2008

  • De Wever, A.; Van der Gucht, K.; Muylaert, K.
  • Aquatic Microbial Ecology, Vol. 50
  • DOI: 10.3354/ame01157

Comparison of the structure and composition of bacterial communities from temperate and tropical freshwater ecosystems
journal, September 2009


Breaking a paradigm: cosmopolitan and abundant freshwater actinobacteria are low GC: Freshwater actinobacteria are low GC
journal, August 2011


Substrate incorporation patterns of bacterioplankton populations in stratified and mixed waters of a humic lake
journal, July 2009


Spatiotemporal distribution and activity patterns of bacteria from three phylogenetic groups in an oligomesotrophic lake
journal, February 2010

  • Salcher, Michaela M.; Pernthaler, Jakob; Posch, Thomas
  • Limnology and Oceanography, Vol. 55, Issue 2
  • DOI: 10.4319/lo.2010.55.2.0846

High-throughput single-cell sequencing identifies photoheterotrophs and chemoautotrophs in freshwater bacterioplankton
journal, June 2011

  • Martinez-Garcia, Manuel; Swan, Brandon K.; Poulton, Nicole J.
  • The ISME Journal, Vol. 6, Issue 1
  • DOI: 10.1038/ismej.2011.84

Metabolic potential of a single cell belonging to one of the most abundant lineages in freshwater bacterioplankton
journal, July 2012

  • Garcia, Sarahi L.; McMahon, Katherine D.; Martinez-Garcia, Manuel
  • The ISME Journal, Vol. 7, Issue 1
  • DOI: 10.1038/ismej.2012.86

In situ substrate preferences of abundant bacterioplankton populations in a prealpine freshwater lake
journal, December 2012

  • Salcher, Michaela M.; Posch, Thomas; Pernthaler, Jakob
  • The ISME Journal, Vol. 7, Issue 5
  • DOI: 10.1038/ismej.2012.162

Key roles for freshwater Actinobacteria revealed by deep metagenomic sequencing
journal, November 2014

  • Ghai, Rohit; Mizuno, Carolina Megumi; Picazo, Antonio
  • Molecular Ecology, Vol. 23, Issue 24
  • DOI: 10.1111/mec.12985

Comparative single-cell genomics reveals potential ecological niches for the freshwater acI Actinobacteria lineage
journal, August 2014

  • Ghylin, Trevor W.; Garcia, Sarahi L.; Moya, Francisco
  • The ISME Journal, Vol. 8, Issue 12
  • DOI: 10.1038/ismej.2014.135

Evaluation of metatranscriptomic protocols and application to the study of freshwater microbial communities: Metatranscriptomics of freshwater communities
journal, June 2014

  • Tsementzi, Despina; Poretsky, Rachel; Rodriguez-R, Luis M.
  • Environmental Microbiology Reports, Vol. 6, Issue 6
  • DOI: 10.1111/1758-2229.12180

Auxotrophy and intrapopulation complementary in the ‘interactome’ of a cultivated freshwater model community
journal, August 2015

  • Garcia, Sarahi L.; Buck, Moritz; McMahon, Katherine D.
  • Molecular Ecology, Vol. 24, Issue 17
  • DOI: 10.1111/mec.13319

A Guide to the Natural History of Freshwater Lake Bacteria
journal, March 2011

  • Newton, R. J.; Jones, S. E.; Eiler, A.
  • Microbiology and Molecular Biology Reviews, Vol. 75, Issue 1
  • DOI: 10.1128/MMBR.00028-10

CheckM: assessing the quality of microbial genomes recovered from isolates, single cells, and metagenomes
journal, May 2015

  • Parks, Donovan H.; Imelfort, Michael; Skennerton, Connor T.
  • Genome Research, Vol. 25, Issue 7
  • DOI: 10.1101/gr.186072.114

OrthoMCL: Identification of Ortholog Groups for Eukaryotic Genomes
journal, September 2003


Actinorhodopsins: proteorhodopsin-like gene sequences found predominantly in non-marine environments
journal, April 2008


Uncoupling of chitinase activity and uptake of hydrolysis products in freshwater bacterioplankton
journal, May 2011


Grazing resistant freshwater bacteria profit from chitin and cell-wall-derived organic carbon: Grazing resistant pelagic scavengers
journal, February 2013

  • Eckert, Ester M.; Baumgartner, Michael; Huber, Iris M.
  • Environmental Microbiology, Vol. 15, Issue 7
  • DOI: 10.1111/1462-2920.12083

Contrasting ability to take up leucine and thymidine among freshwater bacterial groups: implications for bacterial production measurements
journal, January 2010


The Transporter Classification Database
journal, November 2013

  • Saier, Milton H.; Reddy, Vamsee S.; Tamang, Dorjee G.
  • Nucleic Acids Research, Vol. 42, Issue D1
  • DOI: 10.1093/nar/gkt1097

Mixed cultures as model communities: hunting for ubiquitous microorganisms, their partners, and interactions
journal, August 2016


Genome-wide selective sweeps and gene-specific sweeps in natural bacterial populations
journal, January 2016

  • Bendall, Matthew L.; Stevens, Sarah LR; Chan, Leong-Keat
  • The ISME Journal, Vol. 10, Issue 7
  • DOI: 10.1038/ismej.2015.241

PhyloSift: phylogenetic analysis of genomes and metagenomes
journal, January 2014

  • Darling, Aaron E.; Jospin, Guillaume; Lowe, Eric
  • PeerJ, Vol. 2
  • DOI: 10.7717/peerj.243

RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies
journal, January 2014


The large-scale organization of metabolic networks
journal, October 2000

  • Jeong, H.; Tombor, B.; Albert, R.
  • Nature, Vol. 407, Issue 6804
  • DOI: 10.1038/35036627

RASTtk: A modular and extensible implementation of the RAST algorithm for building custom annotation pipelines and annotating batches of genomes
journal, February 2015

  • Brettin, Thomas; Davis, James J.; Disz, Terry
  • Scientific Reports, Vol. 5, Issue 1
  • DOI: 10.1038/srep08365

The SEED and the Rapid Annotation of microbial genomes using Subsystems Technology (RAST)
journal, November 2013

  • Overbeek, Ross; Olson, Robert; Pusch, Gordon D.
  • Nucleic Acids Research, Vol. 42, Issue D1
  • DOI: 10.1093/nar/gkt1226

High-throughput generation, optimization and analysis of genome-scale metabolic models
journal, August 2010

  • Henry, Christopher S.; DeJongh, Matthew; Best, Aaron A.
  • Nature Biotechnology, Vol. 28, Issue 9
  • DOI: 10.1038/nbt.1672

Genomic insights that advance the species definition for prokaryotes
journal, February 2005

  • Konstantinidis, K. T.; Tiedje, J. M.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 7
  • DOI: 10.1073/pnas.0409727102

HTSeq--a Python framework to work with high-throughput sequencing data
journal, September 2014


Mapping and quantifying mammalian transcriptomes by RNA-Seq
journal, May 2008

  • Mortazavi, Ali; Williams, Brian A.; McCue, Kenneth
  • Nature Methods, Vol. 5, Issue 7, p. 621-628
  • DOI: 10.1038/nmeth.1226

PhyloSift: Phylogenetic analysis of genomes and metagenomes
dataset, January 2013


PhyloSift: Phylogenetic analysis of genomes and metagenomes
dataset, January 2013


Works referencing / citing this record:

Linking metagenomics to aquatic microbial ecology and biogeochemical cycles
journal, January 2020

  • Grossart, Hans‐Peter; Massana, Ramon; McMahon, Katherine D.
  • Limnology and Oceanography, Vol. 65, Issue S1
  • DOI: 10.1002/lno.11382

Expansive microbial metabolic versatility and biodiversity in dynamic Guaymas Basin hydrothermal sediments
journal, November 2018


acI Actinobacteria Assemble a Functional Actinorhodopsin with Natively Synthesized Retinal
journal, October 2018

  • Dwulit-Smith, Jeffrey R.; Hamilton, Joshua J.; Stevenson, David M.
  • Applied and Environmental Microbiology, Vol. 84, Issue 24
  • DOI: 10.1128/aem.01678-18

Model Communities Hint at Promiscuous Metabolic Linkages between Ubiquitous Free-Living Freshwater Bacteria
journal, May 2018


Freshwater carbon and nutrient cycles revealed through reconstructed population genomes
journal, January 2018

  • Linz, Alexandra M.; He, Shaomei; Stevens, Sarah L. R.
  • PeerJ, Vol. 6
  • DOI: 10.7717/peerj.6075

Model communities hint at promiscuous metabolic linkages between ubiquitous free-living freshwater bacteria
text, January 2019