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Cap-dependent translation initiation monitored in living cells

Author

Listed:
  • Valentina Gandin

    (Howard Hughes Medical Institute)

  • Brian P. English

    (Howard Hughes Medical Institute)

  • Melanie Freeman

    (Howard Hughes Medical Institute)

  • Louis-Philippe Leroux

    (Institut National de la Recherche Scientifique (INRS)-Centre Armand-Frappier Santé Biotechnologie (CAFSB))

  • Stephan Preibisch

    (Howard Hughes Medical Institute)

  • Deepika Walpita

    (Howard Hughes Medical Institute)

  • Maritza Jaramillo

    (Institut National de la Recherche Scientifique (INRS)-Centre Armand-Frappier Santé Biotechnologie (CAFSB))

  • Robert H. Singer

    (Howard Hughes Medical Institute)

Abstract

mRNA translation is tightly regulated to preserve cellular homeostasis. Despite extensive biochemical, genetic, and structural studies, a detailed understanding of mRNA translation regulation is lacking. Imaging methodologies able to resolve the binding dynamics of translation factors at single-cell and single-mRNA resolution were necessary to fully elucidate regulation of this paramount process. Here live-cell spectroscopy and single-particle tracking were combined to interrogate the binding dynamics of endogenous initiation factors to the 5’cap. The diffusion of initiation factors (IFs) changed markedly upon their association with mRNA. Quantifying their diffusion characteristics revealed the sequence of IFs assembly and disassembly in cell lines and the clustering of translation in neurons. This approach revealed translation regulation at high spatial and temporal resolution that can be applied to the formation of any endogenous complex that results in a measurable shift in diffusion.

Suggested Citation

  • Valentina Gandin & Brian P. English & Melanie Freeman & Louis-Philippe Leroux & Stephan Preibisch & Deepika Walpita & Maritza Jaramillo & Robert H. Singer, 2022. "Cap-dependent translation initiation monitored in living cells," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-34052-8
    DOI: 10.1038/s41467-022-34052-8
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    References listed on IDEAS

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    1. Hannah V. Woodcock & Jessica D. Eley & Delphine Guillotin & Manuela Platé & Carmel B. Nanthakumar & Matteo Martufi & Simon Peace & Gerard Joberty & Daniel Poeckel & Robert B. Good & Adam R. Taylor & N, 2019. "The mTORC1/4E-BP1 axis represents a critical signaling node during fibrogenesis," Nature Communications, Nature, vol. 10(1), pages 1-16, December.
    2. Valentina Gandin & Laia Masvidal & Marie Cargnello & Laszlo Gyenis & Shannon McLaughlan & Yutian Cai & Clara Tenkerian & Masahiro Morita & Preetika Balanathan & Olivier Jean-Jean & Vuk Stambolic & Mat, 2016. "mTORC1 and CK2 coordinate ternary and eIF4F complex assembly," Nature Communications, Nature, vol. 7(1), pages 1-15, September.
    3. Yoshinori Tsukumo & Tommy Alain & Bruno D. Fonseca & Robert Nadon & Nahum Sonenberg, 2016. "Translation control during prolonged mTORC1 inhibition mediated by 4E-BP3," Nature Communications, Nature, vol. 7(1), pages 1-13, September.
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