Dynamics of hunchback translation in real-time and at single-mRNA resolution in the Drosophila embryo
| Autor: | Caroline Hoppe, Daisy J. Vinter, Catherine Sutcliffe, Hilary L. Ashe, Thomas G. Minchington |
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| Rok vydání: | 2021 |
| Předmět: |
Translation
Embryo Nonmammalian animal structures Zygote MS2 SunTag Biology Live cell imaging Translational regulation Morphogenesis Animals Drosophila Proteins Molecular Biology Transcription factor Psychological repression Body Patterning Live imaging Messenger RNA Hunchback Gene Expression Regulation Developmental Embryo Translation (biology) Drosophila embryo Cell biology DNA-Binding Proteins RNA Messenger Stored Protein Biosynthesis Drosophila Transcription Factors Research Article Developmental Biology |
| Zdroj: | Development (Cambridge, England) article-version (VoR) Version of Record |
| ISSN: | 1477-9129 0950-1991 |
| DOI: | 10.1242/dev.196121 |
| Popis: | The Hunchback (Hb) transcription factor is crucial for anterior-posterior patterning of the Drosophila embryo. The maternal hb mRNA acts as a paradigm for translational regulation due to its repression in the posterior of the embryo. However, little is known about the translatability of zygotically transcribed hb mRNAs. Here, we adapt the SunTag system, developed for imaging translation at single-mRNA resolution in tissue culture cells, to the Drosophila embryo to study the translation dynamics of zygotic hb mRNAs. Using single-molecule imaging in fixed and live embryos, we provide evidence for translational repression of zygotic SunTag-hb mRNAs. Whereas the proportion of SunTag-hb mRNAs translated is initially uniform, translation declines from the anterior over time until it becomes restricted to a posterior band in the expression domain. We discuss how regulated hb mRNA translation may help establish the sharp Hb expression boundary, which is a model for precision and noise during developmental patterning. Overall, our data show how use of the SunTag method on fixed and live embryos is a powerful combination for elucidating spatiotemporal regulation of mRNA translation in Drosophila. Summary: Use of the SunTag method to allow single-molecule imaging in fixed and live Drosophila embryos reveals translational repression of zygotic hunchback mRNAs. |
| Databáze: | OpenAIRE |
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