Abstract

Recent functional, proteomic and ribosome profiling studies in eukaryotes have concurrently demonstrated the translation of alternative open reading frames (altORFs) in addition to annotated protein coding sequences (CDSs). We show that a large number of small proteins could in fact be coded by these altORFs. The putative alternative proteins translated from altORFs have orthologs in many species and contain functional domains. Evolutionary analyses indicate that altORFs often show more extreme conservation patterns than their CDSs. Thousands of alternative proteins are detected in proteomic datasets by reanalysis using a database containing predicted alternative proteins. This is illustrated with specific examples, including altMiD51, a 70 amino acid mitochondrial fission-promoting protein encoded in MiD51/Mief1/SMCR7L, a gene encoding an annotated protein promoting mitochondrial fission. Our results suggest that many genes are multicoding genes and code for a large protein and one or several small proteins.

Data availability

The following data sets were generated

Article and author information

Author details

  1. Sondos Samandi

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  2. Annie V Roy

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  3. Vivian Delcourt

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  4. Jean-François Lucier

    Department of Biology, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  5. Jules Gagnon

    Department of Biology, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  6. Maxime C Beaudoin

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  7. Benoît Vanderperre

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  8. Marc-André Breton

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  9. Julie Motard

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  10. Jean-François Jacques

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0465-0313
  11. Mylène Brunelle

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  12. Isabelle Gagnon-Arsenault

    Département de biologie, Université Laval, Quebec, Canada
    Competing interests
    The authors declare that no competing interests exist.
  13. Isabelle Fournier

    Prism INSERM U1192, Université de Lille, Lille, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1096-5044
  14. Aïda Ouangraoua

    Department of Computer Science, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  15. Darel J Hunting

    Department of Nuclear Medicine and Radiobiology, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  16. Alan A Cohen

    Department of Family Medicine, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  17. Christian R Landry

    Département de biologie, Université Laval, Quebec, Canada
    Competing interests
    The authors declare that no competing interests exist.
  18. Michelle S Scott

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    Competing interests
    The authors declare that no competing interests exist.
  19. Xavier Roucou

    Department of Biochemistry, Université de Sherbrooke, Sherbrooke, Canada
    For correspondence
    xavier.roucou@usherbrooke.ca
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9370-5584

Funding

Canadian Institutes of Health Research (MOP-137056)

  • Xavier Roucou

Canada Research Chairs

  • Aïda Ouangraoua
  • Christian R Landry
  • Xavier Roucou

Fonds de Recherche du Québec - Nature et Technologies (2015-PR-181807)

  • Christian R Landry
  • Xavier Roucou

Merck Sharp and Dohme

  • Xavier Roucou

Canadian Institutes of Health Research (MOP-136962)

  • Xavier Roucou

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

Copyright

© 2017, Samandi 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

  • 6,790
    views
  • 1,004
    downloads
  • 97
    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. Sondos Samandi
  2. Annie V Roy
  3. Vivian Delcourt
  4. Jean-François Lucier
  5. Jules Gagnon
  6. Maxime C Beaudoin
  7. Benoît Vanderperre
  8. Marc-André Breton
  9. Julie Motard
  10. Jean-François Jacques
  11. Mylène Brunelle
  12. Isabelle Gagnon-Arsenault
  13. Isabelle Fournier
  14. Aïda Ouangraoua
  15. Darel J Hunting
  16. Alan A Cohen
  17. Christian R Landry
  18. Michelle S Scott
  19. Xavier Roucou
(2017)
Deep transcriptome annotation enables the discovery and functional characterization of cryptic small proteins
eLife 6:e27860.
https://doi.org/10.7554/eLife.27860

Share this article

https://doi.org/10.7554/eLife.27860

Further reading

    1. Biochemistry and Chemical Biology
    Swarang Sachin Pundlik, Alok Barik ... Arvind Ramanathan
    Short Report

    Senescent cells are characterized by multiple features such as increased expression of senescence-associated β-galactosidase activity (SA β-gal) and cell cycle inhibitors such as p21 or p16. They accumulate with tissue damage and dysregulate tissue homeostasis. In the context of skeletal muscle, it is known that agents used for chemotherapy such as Doxorubicin (Doxo) cause buildup of senescent cells, leading to the inhibition of tissue regeneration. Senescent cells influence the neighboring cells via numerous secreted factors which form the senescence-associated secreted phenotype (SASP). Lipids are emerging as a key component of SASP that can control tissue homeostasis. Arachidonic acid-derived lipids have been shown to accumulate within senescent cells, specifically 15d-PGJ2, which is an electrophilic lipid produced by the non-enzymatic dehydration of the prostaglandin PGD2. This study shows that 15d-PGJ2 is also released by Doxo-induced senescent cells as an SASP factor. Treatment of skeletal muscle myoblasts with the conditioned medium from these senescent cells inhibits myoblast fusion during differentiation. Inhibition of L-PTGDS, the enzyme that synthesizes PGD2, diminishes the release of 15d-PGJ2 by senescent cells and restores muscle differentiation. We further show that this lipid post-translationally modifies Cys184 of HRas in C2C12 mouse skeletal myoblasts, causing a reduction in the localization of HRas to the Golgi, increased HRas binding to Ras Binding Domain (RBD) of RAF Kinase (RAF-RBD), and activation of cellular Mitogen Activated Protein (MAP) kinase–Extracellular Signal Regulated Kinase (Erk) signaling (but not the Akt signaling). Mutating C184 of HRas prevents the ability of 15d-PGJ2 to inhibit the differentiation of muscle cells and control the activity of HRas. This work shows that 15d-PGJ2 released from senescent cells could be targeted to restore muscle homeostasis after chemotherapy.

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    Duk-Su Koh, Anastasiia Stratiievska ... Sharona E Gordon
    Tools and Resources

    Ligands such as insulin, epidermal growth factor, platelet-derived growth factor, and nerve growth factor (NGF) initiate signals at the cell membrane by binding to receptor tyrosine kinases (RTKs). Along with G-protein-coupled receptors, RTKs are the main platforms for transducing extracellular signals into intracellular signals. Studying RTK signaling has been a challenge, however, due to the multiple signaling pathways to which RTKs typically are coupled, including MAP/ERK, PLCγ, and Class 1A phosphoinositide 3-kinases (PI3K). The multi-pronged RTK signaling has been a barrier to isolating the effects of any one downstream pathway. Here, we used optogenetic activation of PI3K to decouple its activation from other RTK signaling pathways. In this context, we used genetic code expansion to introduce a click chemistry noncanonical amino acid into the extracellular side of membrane proteins. Applying a cell-impermeant click chemistry fluorophore allowed us to visualize delivery of membrane proteins to the plasma membrane in real time. Using these approaches, we demonstrate that activation of PI3K, without activating other pathways downstream of RTK signaling, is sufficient to traffic the TRPV1 ion channels and insulin receptors to the plasma membrane.