Diverse activities of viral cis-acting RNA regulatory elements revealed using multicolor, long-term, single-cell imaging
| Autor: | Ginger M. Pocock, Kevin W. Eliceiri, Laraine L. Zimdars, Ming Yuan, Nathan M. Sherer, Paul Ahlquist |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Gene Expression Regulation Viral viruses Active Transport Cell Nucleus Biology Regulatory Sequences Nucleic Acid Regulatory Sequences Ribonucleic Acid Bioinformatics Genes env NXF1 03 medical and health sciences Gene expression Murine leukemia virus Methods RNA Messenger RNA Processing Post-Transcriptional Molecular Biology Gene Regulation of gene expression Cell Nucleus Messenger RNA 030102 biochemistry & molecular biology RNA Cell Biology Articles biology.organism_classification 3. Good health Cell biology Molecular Imaging 030104 developmental biology Gene Products rev Regulatory sequence HIV-1 RNA Viral Mason-Pfizer monkey virus Single-Cell Analysis |
| Zdroj: | Molecular Biology of the Cell |
| ISSN: | 1939-4586 |
| Popis: | The regulation of subcellular mRNA localization is a crucial but poorly understood aspect of viral and cellular gene expression. This study demonstrates diverse, striking mRNA subcellular trafficking behaviors and unique gene expression profiles intrinsic to discrete cis-acting RNA structural elements isolated from retroviruses and hepadnaviruses. Cis-acting RNA structural elements govern crucial aspects of viral gene expression. How these structures and other posttranscriptional signals affect RNA trafficking and translation in the context of single cells is poorly understood. Herein we describe a multicolor, long-term (>24 h) imaging strategy for measuring integrated aspects of viral RNA regulatory control in individual cells. We apply this strategy to demonstrate differential mRNA trafficking behaviors governed by RNA elements derived from three retroviruses (HIV-1, murine leukemia virus, and Mason-Pfizer monkey virus), two hepadnaviruses (hepatitis B virus and woodchuck hepatitis virus), and an intron-retaining transcript encoded by the cellular NXF1 gene. Striking behaviors include “burst” RNA nuclear export dynamics regulated by HIV-1’s Rev response element and the viral Rev protein; transient aggregations of RNAs into discrete foci at or near the nuclear membrane triggered by multiple elements; and a novel, pulsiform RNA export activity regulated by the hepadnaviral posttranscriptional regulatory element. We incorporate single-cell tracking and a data-mining algorithm into our approach to obtain RNA element–specific, high-resolution gene expression signatures. Together these imaging assays constitute a tractable, systems-based platform for studying otherwise difficult to access spatiotemporal features of viral and cellular gene regulation. |
| Databáze: | OpenAIRE |
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