1. Cell Biology
  2. Chromosomes and Gene Expression

Biosynthesis of histone messenger RNA employs a specific 3' end endonuclease

  1. Ilaria Pettinati
  2. Pawel Grzechnik
  3. Claudia Ribeiro de Almeida
  4. Jurgen Brem
  5. Michael A McDonough
  6. Somdutta Dhir
  7. Nick J Proudfoot  Is a corresponding author
  8. Christopher J Schofield  Is a corresponding author
  1. University of Oxford, United Kingdom
  2. University of Birmingham, United Kingdom
https://doi.org/10.7554/eLife.39865
Share this article
Cite this article
  1. Ilaria Pettinati
  2. Pawel Grzechnik
  3. Claudia Ribeiro de Almeida
  4. Jurgen Brem
  5. Michael A McDonough
  6. Somdutta Dhir
  7. Nick J Proudfoot
  8. Christopher J Schofield
(2018)
Biosynthesis of histone messenger RNA employs a specific 3' end endonuclease
eLife 7:e39865.
https://doi.org/10.7554/eLife.39865
  2. Download BibTeX
  3. Download .RIS

Abstract

Replication dependent (RD) core histone mRNA produced during S-phase is the only known protein-coding mRNA presenting a 3' stem-loop instead of the otherwise universal polyA tail. A metallo β-lactamase (MBL) fold enzyme, cleavage and polyadenylation specificity factor 73 (CPSF73), is proposed to be the sole endonuclease responsible for 3' end processing of both mRNA classes. We report cellular, genetic, biochemical, substrate selectivity, and crystallographic studies providing evidence that an additional endoribonuclease, MBL domain containing protein 1 (MBLAC1), is selective for 3' processing of RD histone pre-mRNA during the S-phase of the cell cycle. Depletion of MBLAC1 in cells significantly affects cell cycle progression thus identifying MBLAC1 as a new type of S-phase specific cancer target.

Data availability

Diffraction data have been deposited in PDB under the accession code 4V0H.ChrRNA-seq data generated during this study can be accessed at the following link: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=kncnwiswrpapngj&acc=GSE94686

The following data sets were generated

Article and author information

Author details

  1. Ilaria Pettinati

    Department of Chemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.

  2. Pawel Grzechnik

    School of Biosciences, University of Birmingham, Birmingham, United Kingdom
    Competing interests
    No competing interests declared.

  3. Claudia Ribeiro de Almeida

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.

  4. Jurgen Brem

    Department of Chemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.

  5. Michael A McDonough

    Department of Chemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.

  6. Somdutta Dhir

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.

  7. Nick J Proudfoot

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    For correspondence
    nicholas.proudfoot@path.ox.ac.uk
    Competing interests
    Nick J Proudfoot, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8646-3222

  8. Christopher J Schofield

    Department of Chemistry, University of Oxford, Oxford, United Kingdom
    For correspondence
    christopher.schofield@chem.ox.ac.uk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0290-6565

Funding

Wellcome (107928/Z/15/Z)

  • Nick J Proudfoot

Medical Research Council

  • Christopher J Schofield

Wellcome

  • Christopher J Schofield

Cancer Research UK

  • Christopher J Schofield

Biotechnology and Biological Sciences Research Council

  • Christopher J Schofield

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

Copyright

© 2018, Pettinati 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

  • 2,591
    views
  • 387
    downloads
  • 13
    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. Ilaria Pettinati
  2. Pawel Grzechnik
  3. Claudia Ribeiro de Almeida
  4. Jurgen Brem
  5. Michael A McDonough
  6. Somdutta Dhir
  7. Nick J Proudfoot
  8. Christopher J Schofield
(2018)
Biosynthesis of histone messenger RNA employs a specific 3' end endonuclease
eLife 7:e39865.
https://doi.org/10.7554/eLife.39865

Share this article

https://doi.org/10.7554/eLife.39865

Categories and tags

Research organism

Further reading

    1. Cell Biology

    Novel mechanism for tubular injury in nephropathic cystinosis

    Swastika Sur, Maggie Kerwin ... Minnie M Sarwal
    Research Article

    Understanding the unique susceptibility of the human kidney to pH dysfunction and injury in cystinosis is paramount to developing new therapies to preserve renal function. Renal proximal tubular epithelial cells (RPTECs) and fibroblasts isolated from patients with cystinosis were transcriptionally profiled. Lysosomal fractionation, immunoblotting, confocal microscopy, intracellular pH, TEM, and mitochondrial stress test were performed for validation. CRISPR, CTNS -/- RPTECs were generated. Alterations in cell stress, pH, autophagic turnover, and mitochondrial energetics highlighted key changes in the V-ATPases in patient-derived and CTNS-/- RPTECs. ATP6V0A1 was significantly downregulated in cystinosis and highly co-regulated with loss of CTNS. Correction of ATP6V0A1 rescued cell stress and mitochondrial function. Treatment of CTNS -/- RPTECs with antioxidants ATX induced ATP6V0A1 expression and improved autophagosome turnover and mitochondrial integrity. Our exploratory transcriptional and in vitro cellular and functional studies confirm that loss of Cystinosin in RPTECs, results in a reduction in ATP6V0A1 expression, with changes in intracellular pH, mitochondrial integrity, mitochondrial function, and autophagosome-lysosome clearance. The novel findings are ATP6V0A1’s role in cystinosis-associated renal pathology and among other antioxidants, ATX specifically upregulated ATP6V0A1, improved autophagosome turnover or reduced autophagy and mitochondrial integrity. This is a pilot study highlighting a novel mechanism of tubular injury in cystinosis.

    1. Cell Biology
    2. Developmental Biology

    Ovarian Biology: Revisiting the rete ovarii

    Yan Zhang, Hua Zhang
    Insight

    Long thought to have little relevance to ovarian physiology, the rete ovarii may have a role in follicular dynamics and reproductive health.

    1. Cell Biology
    2. Developmental Biology

    Rediscovering the rete ovarii, a secreting auxiliary structure to the ovary

    Dilara N Anbarci, Jennifer McKey ... Blanche Capel
    Research Article

    The rete ovarii (RO) is an appendage of the ovary that has been given little attention. Although the RO appears in drawings of the ovary in early versions of Gray’s Anatomy, it disappeared from recent textbooks, and is often dismissed as a functionless vestige in the adult ovary. Using PAX8 immunostaining and confocal microscopy, we characterized the fetal development of the RO in the context of the mouse ovary. The RO consists of three distinct regions that persist in adult life, the intraovarian rete (IOR), the extraovarian rete (EOR), and the connecting rete (CR). While the cells of the IOR appear to form solid cords within the ovary, the EOR rapidly develops into a convoluted tubular epithelium ending in a distal dilated tip. Cells of the EOR are ciliated and exhibit cellular trafficking capabilities. The CR, connecting the EOR to the IOR, gradually acquires tubular epithelial characteristics by birth. Using microinjections into the distal dilated tip of the EOR, we found that luminal contents flow toward the ovary. Mass spectrometry revealed that the EOR lumen contains secreted proteins potentially important for ovarian function. We show that the cells of the EOR are closely associated with vasculature and macrophages, and are contacted by neuronal projections, consistent with a role as a sensory appendage of the ovary. The direct proximity of the RO to the ovary and its integration with the extraovarian landscape suggest that it plays an important role in ovary development and homeostasis.