SLC38A2 provides proline to fulfil unique synthetic demands arising during osteoblast differentiation and bone formation

  1. Leyao Shen
  2. Yilin Yu
  3. Yunji Zhou
  4. Shondra M Pruett-Miller
  5. Guo-Fang Zhang
  6. Courtney M Karner  Is a corresponding author
  1. University of Texas Southwestern Medical Center, United States
  2. Duke University, United States
  3. St Jude Children's Research Hospital, United States
  4. Duke University Medical Center, United States

Abstract

Cellular differentiation is associated with the acquisition of a unique protein signature which is essential to attain the ultimate cellular function and activity of the differentiated cell. This is predicted to result in unique biosynthetic demands that arise during differentiation. Using a bioinformatic approach, we discovered osteoblast differentiation is associated with increased demand for the amino acid proline. When compared to other differentiated cells, osteoblast-associated proteins including RUNX2, OSX, OCN and COL1A1 are significantly enriched in proline. Using a genetic and metabolomic approach, we demonstrate that the neutral amino acid transporter SLC38A2 acts cell autonomously to provide proline to facilitate the efficient synthesis of proline-rich osteoblast proteins. Genetic ablation of SLC38A2 in osteoblasts limits both osteoblast differentiation and bone formation in mice. Mechanistically, proline is primarily incorporated into nascent protein with little metabolism observed. Collectively, these data highlight a requirement for proline in fulfilling the unique biosynthetic requirements that arise during osteoblast differentiation and bone formation.

Data availability

All data generated or analyzed during this study are included in this submission and the supporting files. Source data files are included for all western blot images and excel spreadsheets are included for the RNAseq and metabolic tracing experiments in figures 1 and 2.

Article and author information

Author details

  1. Leyao Shen

    Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Yilin Yu

    Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Yunji Zhou

    Department of Biostatistics and Bioinformatics, Duke University, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Shondra M Pruett-Miller

    Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3793-585X
  5. Guo-Fang Zhang

    Sarah W Stedman Nutrition and Metabolism Center, Duke University Medical Center, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Courtney M Karner

    Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, United States
    For correspondence
    courtney.karner@utsouthwestern.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0387-4486

Funding

National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR071967)

  • Courtney M Karner

National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR076325)

  • Courtney M Karner

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

Reviewing Editor

  1. Ernestina Schipani, University of Pennsylvania, United States

Ethics

Animal experimentation: All mouse procedures were approved by the Animal Studies Committees at Duke University first and then the University of Texas Southwestern Medical Center at Dallas (Animal Protocol 2020-102999).

Version history

  1. Received: January 11, 2022
  2. Preprint posted: January 12, 2022 (view preprint)
  3. Accepted: March 8, 2022
  4. Accepted Manuscript published: March 9, 2022 (version 1)
  5. Accepted Manuscript updated: March 15, 2022 (version 2)
  6. Version of Record published: April 13, 2022 (version 3)

Copyright

© 2022, Shen 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.

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  1. Leyao Shen
  2. Yilin Yu
  3. Yunji Zhou
  4. Shondra M Pruett-Miller
  5. Guo-Fang Zhang
  6. Courtney M Karner
(2022)
SLC38A2 provides proline to fulfil unique synthetic demands arising during osteoblast differentiation and bone formation
eLife 11:e76963.
https://doi.org/10.7554/eLife.76963

Share this article

https://doi.org/10.7554/eLife.76963

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