DAZL mediates a broad translational program regulating expansion and differentiation of spermatogonial progenitors

Abstract

Fertility across metazoa requires the germline-specific DAZ family of RNA-binding proteins. Here we examine whether DAZL directly regulates progenitor spermatogonia using a conditional genetic mouse model and in vivo biochemical approaches combined with chemical synchronization of spermatogenesis. We find that the absence of Dazl impairs both expansion and differentiation of the spermatogonial progenitor population. In undifferentiated spermatogonia, DAZL binds the 3' UTRs of ~2,500 protein-coding genes. Some targets are known regulators of spermatogonial proliferation and differentiation while others are broadly expressed, dosage-sensitive factors that control transcription and RNA metabolism. DAZL binds 3' UTR sites conserved across vertebrates at a UGUU(U/A) motif. By assessing ribosome occupancy in undifferentiated spermatogonia, we find that DAZL increases translation of its targets. In total, DAZL orchestrates a broad translational program that amplifies protein levels of key spermatogonial and gene regulatory factors to promote the expansion and differentiation of progenitor spermatogonia.

Data availability

All sequencing data generated in this study are available at NCBI Gene Expression Omnibus accession number GSE145177

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Maria M Mikedis

    Whitehead Institute, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Yuting Fan

    Whitehead Institute, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Peter K Nicholls

    Whitehead Institute, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5540-442X
  4. Tsutomu Endo

    Whitehead Institute, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Emily K Jackson

    Whitehead Institute, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Sarah A Cobb

    Whitehead Institute, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Dirk G de Rooij

    Whitehead Institute, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3932-4419
  8. David C Page

    Whitehead Institute, Cambridge, United States
    For correspondence
    dcpage@wi.mit.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9920-3411

Funding

Howard Hughes Medical Institute (Page laboratory)

  • David C Page

Lalor Foundation (Postdoctoral fellowship)

  • Maria M Mikedis

Eunice Kennedy Shriver National Institute of Child Health and Human Development (F32HD093391)

  • Maria M Mikedis

National Natural Science Foundation of China (81471507)

  • Yuting Fan

National Key Research and Development Program of China (2017YFC1001600)

  • Yuting Fan

Hope Funds for Cancer Research (HFCR-15-06-06)

  • Peter K Nicholls

National Health and Medical Research Council (GNT1053776)

  • Peter K Nicholls

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

Ethics

Animal experimentation: All experiments involving mice were performed in accordance with the guidelines of the Massachusetts Institute of Technology (MIT) Division of Comparative Medicine, which is overseen by MIT's Institutional Animal Care and Use Committee (IACUC). The animal care program at MIT/Whitehead Institute is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International (AAALAC), and meets or exceeds the standards of AAALAC as detailed in the Guide for the Care and Use of Laboratory Animals. The MIT IACUC approved this research (no. 0617-059-20).

Reviewing Editor

  1. Moira K O'Bryan, Monash University, Australia

Publication history

  1. Received: March 2, 2020
  2. Accepted: July 20, 2020
  3. Accepted Manuscript published: July 20, 2020 (version 1)
  4. Version of Record published: August 24, 2020 (version 2)

Copyright

© 2020, Mikedis 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

  • 2,042
    Page views
  • 355
    Downloads
  • 20
    Citations

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

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. Maria M Mikedis
  2. Yuting Fan
  3. Peter K Nicholls
  4. Tsutomu Endo
  5. Emily K Jackson
  6. Sarah A Cobb
  7. Dirk G de Rooij
  8. David C Page
(2020)
DAZL mediates a broad translational program regulating expansion and differentiation of spermatogonial progenitors
eLife 9:e56523.
https://doi.org/10.7554/eLife.56523

Further reading

    1. Developmental Biology
    Tsz Long Chu, Peikai Chen ... Kathryn Song Eng Cheah
    Research Article Updated

    Bone homeostasis is regulated by hormones such as parathyroid hormone (PTH). While PTH can stimulate osteo-progenitor expansion and bone synthesis, how the PTH-signaling intensity in progenitors is controlled is unclear. Endochondral bone osteoblasts arise from perichondrium-derived osteoprogenitors and hypertrophic chondrocytes (HC). We found, via single-cell transcriptomics, that HC-descendent cells activate membrane-type 1 metalloproteinase 14 (MMP14) and the PTH pathway as they transition to osteoblasts in neonatal and adult mice. Unlike Mmp14 global knockouts, postnatal day 10 (p10) HC lineage-specific Mmp14 null mutants (Mmp14ΔHC) produce more bone. Mechanistically, MMP14 cleaves the extracellular domain of PTH1R, dampening PTH signaling, and consistent with the implied regulatory role, in Mmp14ΔHC mutants, PTH signaling is enhanced. We found that HC-derived osteoblasts contribute ~50% of osteogenesis promoted by treatment with PTH 1–34, and this response was amplified in Mmp14ΔHC. MMP14 control of PTH signaling likely applies also to both HC- and non-HC-derived osteoblasts because their transcriptomes are highly similar. Our study identifies a novel paradigm of MMP14 activity-mediated modulation of PTH signaling in the osteoblast lineage, contributing new insights into bone metabolism with therapeutic significance for bone-wasting diseases.

    1. Biochemistry and Chemical Biology
    2. Developmental Biology
    Zengdi Zhang, Zan Huang ... Hai-Bin Ruan
    Research Article Updated

    In mammals, interactions between the bone marrow (BM) stroma and hematopoietic progenitors contribute to bone-BM homeostasis. Perinatal bone growth and ossification provide a microenvironment for the transition to definitive hematopoiesis; however, mechanisms and interactions orchestrating the development of skeletal and hematopoietic systems remain largely unknown. Here, we establish intracellular O-linked β-N-acetylglucosamine (O-GlcNAc) modification as a posttranslational switch that dictates the differentiation fate and niche function of early BM stromal cells (BMSCs). By modifying and activating RUNX2, O-GlcNAcylation promotes osteogenic differentiation of BMSCs and stromal IL-7 expression to support lymphopoiesis. In contrast, C/EBPβ-dependent marrow adipogenesis and expression of myelopoietic stem cell factor (SCF) is inhibited by O-GlcNAcylation. Ablating O-GlcNAc transferase (OGT) in BMSCs leads to impaired bone formation, increased marrow adiposity, as well as defective B-cell lymphopoiesis and myeloid overproduction in mice. Thus, the balance of osteogenic and adipogenic differentiation of BMSCs is determined by reciprocal O-GlcNAc regulation of transcription factors, which simultaneously shapes the hematopoietic niche.