On the flexibility of the cellular amination network in E. coli

  1. Helena Schulz-Mirbach
  2. Alexandra Müller
  3. Tong Wu
  4. Pascal Pfister
  5. Selçuk Aslan
  6. Lennart Schada von Borzyskowski
  7. Tobias J Erb
  8. Arren Bar-Even
  9. Steffen N Lindner  Is a corresponding author
  1. Max Planck Institute of Molecular Plant Physiology, Germany
  2. Max Planck Institute for Terrestrial Microbiology, Germany

Abstract

Ammonium (NH4+) is essential to generate the nitrogenous building blocks of life. It gets assimilated via the canonical biosynthetic routes to glutamate and is further distributed throughout metabolism via a network of transaminases. To study the flexibility of this network, we constructed an Escherichia coli glutamate auxotrophic strain. This strain allowed us to systematically study which amino acids serve as amine sources and found that several amino acids complement the auxotrophy, either by producing glutamate via transamination reactions or by their conversion to glutamate. In this network, we identified aspartate transaminase AspC as a major connector between many amino acids and glutamate. Additionally, we extended the transaminase network by the amino acids β-alanine, alanine, glycine, and serine as new amine sources and identified d-amino acid dehydrogenase (DadA) as an intracellular amino acid sink removing substrates from transaminase reactions. Finally, ammonium assimilation routes producing aspartate or leucine were introduced. Our study reveals the high flexibility of the cellular amination network, both in terms of transaminase promiscuity and adaptability to new connections and ammonium entry points.

Data availability

sequencing data has been deposited at Dryad

The following data sets were generated

Article and author information

Author details

  1. Helena Schulz-Mirbach

    Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5376-9185
  2. Alexandra Müller

    Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Tong Wu

    Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Pascal Pfister

    Max Planck Institute for Terrestrial Microbiology, Marburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Selçuk Aslan

    Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Lennart Schada von Borzyskowski

    Max Planck Institute for Terrestrial Microbiology, Marburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Tobias J Erb

    Max Planck Institute for Terrestrial Microbiology, Marburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Arren Bar-Even

    Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1039-4328
  9. Steffen N Lindner

    Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
    For correspondence
    Lindner@mpimp-golm.mpg.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3226-3043

Funding

Bundesministerium für Bildung und Forschung (031B0825B)

  • Steffen N Lindner

Deutsche Forschungsgemeinschaft (SFB987)

  • Tobias J Erb

Max Planck Institute of Molecular Plant Physiology (open access funding)

  • Helena Schulz-Mirbach
  • Alexandra Müller
  • Tong Wu
  • Selçuk Aslan
  • Arren Bar-Even
  • Steffen N Lindner

Max Planck Institute for Terrestrial Microbiology (open access funding)

  • Pascal Pfister

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Ahmad S Khalil, Boston University, United States

Version history

  1. Preprint posted: January 27, 2022 (view preprint)
  2. Received: February 1, 2022
  3. Accepted: July 22, 2022
  4. Accepted Manuscript published: July 25, 2022 (version 1)
  5. Version of Record published: September 1, 2022 (version 2)

Copyright

© 2022, Schulz-Mirbach et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,608
    views
  • 452
    downloads
  • 3
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Helena Schulz-Mirbach
  2. Alexandra Müller
  3. Tong Wu
  4. Pascal Pfister
  5. Selçuk Aslan
  6. Lennart Schada von Borzyskowski
  7. Tobias J Erb
  8. Arren Bar-Even
  9. Steffen N Lindner
(2022)
On the flexibility of the cellular amination network in E. coli
eLife 11:e77492.
https://doi.org/10.7554/eLife.77492

Share this article

https://doi.org/10.7554/eLife.77492

Further reading

    1. Cell Biology
    2. Chromosomes and Gene Expression
    Lucie Crhak Khaitova, Pavlina Mikulkova ... Karel Riha
    Research Article

    Heat stress is a major threat to global crop production, and understanding its impact on plant fertility is crucial for developing climate-resilient crops. Despite the known negative effects of heat stress on plant reproduction, the underlying molecular mechanisms remain poorly understood. Here, we investigated the impact of elevated temperature on centromere structure and chromosome segregation during meiosis in Arabidopsis thaliana. Consistent with previous studies, heat stress leads to a decline in fertility and micronuclei formation in pollen mother cells. Our results reveal that elevated temperature causes a decrease in the amount of centromeric histone and the kinetochore protein BMF1 at meiotic centromeres with increasing temperature. Furthermore, we show that heat stress increases the duration of meiotic divisions and prolongs the activity of the spindle assembly checkpoint during meiosis I, indicating an impaired efficiency of the kinetochore attachments to spindle microtubules. Our analysis of mutants with reduced levels of centromeric histone suggests that weakened centromeres sensitize plants to elevated temperature, resulting in meiotic defects and reduced fertility even at moderate temperatures. These results indicate that the structure and functionality of meiotic centromeres in Arabidopsis are highly sensitive to heat stress, and suggest that centromeres and kinetochores may represent a critical bottleneck in plant adaptation to increasing temperatures.

    1. Cell Biology
    Wan-ping Yang, Mei-qi Li ... Qian-qian Luo
    Research Article

    High-altitude polycythemia (HAPC) affects individuals living at high altitudes, characterized by increased red blood cells (RBCs) production in response to hypoxic conditions. The exact mechanisms behind HAPC are not fully understood. We utilized a mouse model exposed to hypobaric hypoxia (HH), replicating the environmental conditions experienced at 6000 m above sea level, coupled with in vitro analysis of primary splenic macrophages under 1% O2 to investigate these mechanisms. Our findings indicate that HH significantly boosts erythropoiesis, leading to erythrocytosis and splenic changes, including initial contraction to splenomegaly over 14 days. A notable decrease in red pulp macrophages (RPMs) in the spleen, essential for RBCs processing, was observed, correlating with increased iron release and signs of ferroptosis. Prolonged exposure to hypoxia further exacerbated these effects, mirrored in human peripheral blood mononuclear cells. Single-cell sequencing showed a marked reduction in macrophage populations, affecting the spleen’s ability to clear RBCs and contributing to splenomegaly. Our findings suggest splenic ferroptosis contributes to decreased RPMs, affecting erythrophagocytosis and potentially fostering continuous RBCs production in HAPC. These insights could guide the development of targeted therapies for HAPC, emphasizing the importance of splenic macrophages in disease pathology.