1. Biochemistry and Chemical Biology
  2. Cell Biology
Download icon

Identification of a transporter complex responsible for the cytosolic entry of nitrogen-containing-bisphosphonates

  1. Zhou Yu
  2. Lauren E Surface
  3. Chong Yon Park
  4. Max A Horlbeck
  5. Gregory A Wyant
  6. Monther Abu-Remaileh
  7. Timothy R Peterson
  8. David M Sabatini
  9. Jonathan S Weissman
  10. Erin K O'Shea  Is a corresponding author
  1. Harvard University, United States
  2. University of California, San Francisco, United States
  3. Whitehead Institute for Biomedical Research, United States
  4. Washington University School of Medicine, United States
Short Report
  • Cited 19
  • Views 2,830
  • Annotations
Cite this article as: eLife 2018;7:e36620 doi: 10.7554/eLife.36620

Abstract

Nitrogen-containing-bisphosphonates (N-BPs) are a class of drugs widely prescribed to treat osteoporosis and other bone-related diseases. Although previous studies have established that N-BPs function by inhibiting the mevalonate pathway in osteoclasts, the mechanism by which N-BPs enter the cytosol from the extracellular space to reach their molecular target is not understood. Here we implemented a CRISPRi-mediated genome-wide screen and identified SLC37A3 (solute carrier family 37 member A3) as a gene required for the action of N-BPs in mammalian cells. We observed that SLC37A3 forms a complex with ATRAID (all-trans retinoic acid-induced differentiation factor), a previously identified genetic target of N-BPs. SLC37A3 and ATRAID localize to lysosomes and are required for releasing N-BP molecules that have trafficked to lysosomes through fluid-phase endocytosis into the cytosol. Our results elucidate the route by which N-BPs are delivered to their molecular target, addressing a key aspect of the mechanism of action of N-BPs that may have significant clinical relevance.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for the two CRISPRi screens shown in Figure 1 and its figure supplement.

The following data sets were generated

Article and author information

Author details

  1. Zhou Yu

    Department of Molecular and Cellular Biology, Harvard University, Cambridge, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3213-4583

  2. Lauren E Surface

    Department of Molecular and Cellular Biology, Harvard University, Cambridge, United States
    Competing interests
    No competing interests declared.

  3. Chong Yon Park

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.

  4. Max A Horlbeck

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3875-871X

  5. Gregory A Wyant

    Whitehead Institute for Biomedical Research, Cambridge, United States
    Competing interests
    No competing interests declared.

  6. Monther Abu-Remaileh

    Whitehead Institute for Biomedical Research, Cambridge, United States
    Competing interests
    No competing interests declared.

  7. Timothy R Peterson

    Division of Bone and Mineral Diseases, Washington University School of Medicine, St. Louis, United States
    Competing interests
    No competing interests declared.

  8. David M Sabatini

    Whitehead Institute for Biomedical Research, Cambridge, United States
    Competing interests
    No competing interests declared.

  9. Jonathan S Weissman

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2445-670X

  10. Erin K O'Shea

    Department of Molecular and Cellular Biology, Harvard University, Cambridge, United States
    For correspondence
    osheae@hhmi.org
    Competing interests
    Erin K O'Shea, Chief Scientific Officer and a Vice President at the Howard Hughes Medical Institute, one of the three founding funders of eLife..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2649-1018

Funding

Howard Hughes Medical Institute

  • Zhou Yu
  • Lauren E Surface
  • Chong Yon Park
  • Max A Horlbeck
  • Gregory A Wyant
  • Monther Abu-Remaileh
  • David M Sabatini
  • Jonathan S Weissman
  • Erin K O'Shea

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

Reviewing Editor

  1. Philip A Cole, Harvard Medical School, United States

Publication history

  1. Received: March 13, 2018
  2. Accepted: May 9, 2018
  3. Accepted Manuscript published: May 10, 2018 (version 1)
  4. Version of Record published: June 27, 2018 (version 2)

Copyright

© 2018, Yu 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

  • 2,830
    Page views
  • 503
    Downloads
  • 19
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

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)

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

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

Categories and tags

Research organisms

    1. Related to

    Bone-Targeting Drugs: From vesicle to cytosol

    Michael J Rogers, Marcia A Munoz
  2. Further reading

Further reading

    1. Biochemistry and Chemical Biology
    2. Cell Biology

    Bone-Targeting Drugs: From vesicle to cytosol

    Michael J Rogers, Marcia A Munoz
    Insight

    Drugs called bisphosphonates are used to treat a range of bone diseases, but how do they reach the enzymes that are their target?

    1. Biochemistry and Chemical Biology

    Myopalladin knockout mice develop cardiac dilation and show a maladaptive response to mechanical pressure overload

    Maria Carmela Filomena et al.
    Research Article

    Myopalladin (MYPN) is a striated muscle-specific immunoglobulin domain-containing protein located in the sarcomeric Z-line and I-band. MYPN gene mutations are causative for dilated (DCM), hypertrophic and restrictive cardiomyopathy. In a yeast two-hybrid screening, MYPN was found to bind to titin in the Z-line, which was confirmed by microscale thermophoresis. Cardiac analyses of MYPN knockout (MKO) mice showed the development of mild cardiac dilation and systolic dysfunction, associated with decreased myofibrillar isometric tension generation and increased resting tension at longer sarcomere lengths. MKO mice exhibited a normal hypertrophic response to transaortic constriction (TAC), but rapidly developed severe cardiac dilation and systolic dysfunction, associated with fibrosis, increased fetal gene expression, higher intercalated disc fold amplitude, decreased calsequestrin-2 protein levels, and increased desmoplakin and SORBS2 protein levels. Cardiomyocyte analyses showed delayed Ca2+ release and reuptake in unstressed MKO mice as well as reduced Ca2+ spark amplitude post-TAC, suggesting that altered Ca2+ handling may contribute to the development of DCM in MKO mice.

    1. Biochemistry and Chemical Biology

    Witnessing the structural evolution of an RNA enzyme

    Xavier Portillo et al.
    Research Article Updated

    An RNA polymerase ribozyme that has been the subject of extensive directed evolution efforts has attained the ability to synthesize complex functional RNAs, including a full-length copy of its own evolutionary ancestor. During the course of evolution, the catalytic core of the ribozyme has undergone a major structural rearrangement, resulting in a novel tertiary structural element that lies in close proximity to the active site. Through a combination of site-directed mutagenesis, structural probing, and deep sequencing analysis, the trajectory of evolution was seen to involve the progressive stabilization of the new structure, which provides the basis for improved catalytic activity of the ribozyme. Multiple paths to the new structure were explored by the evolving population, converging upon a common solution. Tertiary structural remodeling of RNA is known to occur in nature, as evidenced by the phylogenetic analysis of extant organisms, but this type of structural innovation had not previously been observed in an experimental setting. Despite prior speculation that the catalytic core of the ribozyme had become trapped in a narrow local fitness optimum, the evolving population has broken through to a new fitness locale, raising the possibility that further improvement of polymerase activity may be achievable.