Maintenance of Miranda Localization in Drosophila Neuroblasts Involves Interaction with the Cognate mRNA

Anne Ramat; Matthew Hannaford; Jens Januschke

doi:10.1016/j.cub.2017.06.016

Maintenance of Miranda Localization in Drosophila Neuroblasts Involves Interaction with the Cognate mRNA

Anne Ramat, Matthew Hannaford, Jens Januschke (Lead / Corresponding author)

Research output: Contribution to journal › Article › peer-review

20 Citations (Scopus)

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Abstract

How cells position their proteins is a key problem in cell biology. Targeting mRNAs to distinct regions of the cytoplasm contributes to protein localization by providing local control over translation. Here we reveal that an interdependence of a protein and cognate mRNA maintains asymmetric protein distribution in mitotic Drosophila neural stem cells. We tagged endogenous mRNA or protein products of the gene miranda that is required for fate determination with GFP. We find that the mRNA localizes like the protein it encodes in a basal crescent in mitosis. We then used GFP-specific nanobodies fused to localization domains to alter the subcellular distribution of the GFP-tagged mRNA or protein. Altering the localization of the mRNA resulted in mislocalization of the protein and vice versa. Protein localization defects caused by mislocalization of the cognate mRNA were rescued by introducing untagged mRNA coding for mutant non-localizable protein. Therefore by combining the MS2 system and subcellular nanobody expression we uncovered that maintenance of Mira asymmetric localization requires interaction with the cognate mRNA.

Original language	English
Pages (from-to)	2101-2015
Number of pages	16
Journal	Current Biology
Volume	27
Issue number	14
Early online date	6 Jul 2017
DOIs	https://doi.org/10.1016/j.cub.2017.06.016
Publication status	Published - 6 Jul 2017

Keywords

Drosophila
Asymmetric cell division
mRNA localization
Nanobody
Neuroblasts
Polarity

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10.1016/j.cub.2017.06.016Licence: CC BY

Final Published Version
©2017 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Final published version, 5.35 MBLicence: CC BY

Recycling Polarity - Mechanisms Controlling Stem Cell Polarity in Consecutive Divisions in the Developing Drosophila Central Nervous System (Sir Henry Dale Fellowship)
Januschke, J. (Investigator) & Storey, K. (Investigator)
1/02/13 → 30/11/21
Project: Research

Establishing and Maintaining Cortical Asymmetry in Drosophila Neural Stem Cells
Hannaford, M. (Author), Januschke, J. (Supervisor), 2017
Student thesis: Doctoral Thesis › Doctor of Philosophy

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Januschke, Jens
- Molecular Cell and Developmental Biology - Professor (Teaching and Research) of Development Cell Biology
Person: Academic

Cite this

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title = "Maintenance of Miranda Localization in Drosophila Neuroblasts Involves Interaction with the Cognate mRNA",

abstract = "How cells position their proteins is a key problem in cell biology. Targeting mRNAs to distinct regions of the cytoplasm contributes to protein localization by providing local control over translation. Here we reveal that an interdependence of a protein and cognate mRNA maintains asymmetric protein distribution in mitotic Drosophila neural stem cells. We tagged endogenous mRNA or protein products of the gene miranda that is required for fate determination with GFP. We find that the mRNA localizes like the protein it encodes in a basal crescent in mitosis. We then used GFP-specific nanobodies fused to localization domains to alter the subcellular distribution of the GFP-tagged mRNA or protein. Altering the localization of the mRNA resulted in mislocalization of the protein and vice versa. Protein localization defects caused by mislocalization of the cognate mRNA were rescued by introducing untagged mRNA coding for mutant non-localizable protein. Therefore by combining the MS2 system and subcellular nanobody expression we uncovered that maintenance of Mira asymmetric localization requires interaction with the cognate mRNA.",

keywords = "Drosophila , Asymmetric cell division , mRNA localization , Nanobody , Neuroblasts , Polarity",

author = "Anne Ramat and Matthew Hannaford and Jens Januschke",

note = "A.R. is supported by the Wellcome Institutional Strategic Funds Grant 097818/Z/11/Z. M.R.H is supported by an MRC studentship funded by these grants: G1000386/1, MR/J50046X/1, MR/K500896/1, MR/K501384/1. Work in JJ{\textquoteright}s lab is supported by a Sir Henry Dale fellowship from Wellcome and the Royal Society 100031Z/12/Z. The tissue imaging facility is supported by the grant WT101468 from Wellcome.",

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N1 - A.R. is supported by the Wellcome Institutional Strategic Funds Grant 097818/Z/11/Z. M.R.H is supported by an MRC studentship funded by these grants: G1000386/1, MR/J50046X/1, MR/K500896/1, MR/K501384/1. Work in JJ’s lab is supported by a Sir Henry Dale fellowship from Wellcome and the Royal Society 100031Z/12/Z. The tissue imaging facility is supported by the grant WT101468 from Wellcome.

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N2 - How cells position their proteins is a key problem in cell biology. Targeting mRNAs to distinct regions of the cytoplasm contributes to protein localization by providing local control over translation. Here we reveal that an interdependence of a protein and cognate mRNA maintains asymmetric protein distribution in mitotic Drosophila neural stem cells. We tagged endogenous mRNA or protein products of the gene miranda that is required for fate determination with GFP. We find that the mRNA localizes like the protein it encodes in a basal crescent in mitosis. We then used GFP-specific nanobodies fused to localization domains to alter the subcellular distribution of the GFP-tagged mRNA or protein. Altering the localization of the mRNA resulted in mislocalization of the protein and vice versa. Protein localization defects caused by mislocalization of the cognate mRNA were rescued by introducing untagged mRNA coding for mutant non-localizable protein. Therefore by combining the MS2 system and subcellular nanobody expression we uncovered that maintenance of Mira asymmetric localization requires interaction with the cognate mRNA.

AB - How cells position their proteins is a key problem in cell biology. Targeting mRNAs to distinct regions of the cytoplasm contributes to protein localization by providing local control over translation. Here we reveal that an interdependence of a protein and cognate mRNA maintains asymmetric protein distribution in mitotic Drosophila neural stem cells. We tagged endogenous mRNA or protein products of the gene miranda that is required for fate determination with GFP. We find that the mRNA localizes like the protein it encodes in a basal crescent in mitosis. We then used GFP-specific nanobodies fused to localization domains to alter the subcellular distribution of the GFP-tagged mRNA or protein. Altering the localization of the mRNA resulted in mislocalization of the protein and vice versa. Protein localization defects caused by mislocalization of the cognate mRNA were rescued by introducing untagged mRNA coding for mutant non-localizable protein. Therefore by combining the MS2 system and subcellular nanobody expression we uncovered that maintenance of Mira asymmetric localization requires interaction with the cognate mRNA.

KW - Drosophila

KW - Asymmetric cell division

KW - mRNA localization

KW - Nanobody

KW - Neuroblasts

KW - Polarity

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DO - 10.1016/j.cub.2017.06.016

M3 - Article

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SN - 0960-9822

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JO - Current Biology

JF - Current Biology

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ER -

Maintenance of Miranda Localization in Drosophila Neuroblasts Involves Interaction with the Cognate mRNA

Abstract

Keywords

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Recycling Polarity - Mechanisms Controlling Stem Cell Polarity in Consecutive Divisions in the Developing Drosophila Central Nervous System (Sir Henry Dale Fellowship)

Establishing and Maintaining Cortical Asymmetry in Drosophila Neural Stem Cells

Januschke, Jens

Cite this

Maintenance of Miranda Localization in Drosophila Neuroblasts Involves Interaction with the Cognate mRNA

Abstract

Keywords

Access to Document

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Projects

Recycling Polarity - Mechanisms Controlling Stem Cell Polarity in Consecutive Divisions in the Developing Drosophila Central Nervous System (Sir Henry Dale Fellowship)

Student theses

Establishing and Maintaining Cortical Asymmetry in Drosophila Neural Stem Cells

Profiles

Januschke, Jens

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