Single-Molecule Sensitivity RNA FISH Analysis of Influenza Virus Genome Trafficking

2018 ◽  
pp. 195-211
Author(s):  
Yi-ying Chou ◽  
Timothée Lionnet
Keyword(s):  
Influenza Virus ◽  
Single Molecule ◽  
Virus Genome ◽  
Fish Analysis ◽  
Rna Fish

scholarly journals Transcription factor residence time dominates over concentration in transcription activation

2020 ◽  
Author(s):  
Achim P. Popp ◽  
Johannes Hettich ◽  
J. Christof M. Gebhardt
Keyword(s):  
Transcription Factor ◽  
Residence Time ◽  
Single Molecule ◽  
Search Time ◽  
Mrna Synthesis ◽  
Burst Frequency ◽  
Quantitative Measurements ◽  
Rna Fish ◽  
Transition Times ◽  
Basic Course

Transcription is a vital process activated by transcription factor (TF) binding. The active gene releases a burst of transcripts before turning inactive again. While the basic course of transcription is well understood, it is unclear how binding of a TF affects the frequency, duration and size of a transcriptional burst. We systematically varied the residence time and concentration of a synthetic TF and characterized the transcription of a reporter gene by combining single molecule imaging, single molecule RNA-FISH, live transcript visualisation and analysis with a novel algorithm, Burst Inference from mRNA Distributions (BIRD). For this well-defined system, we found that TF binding solely affected burst frequency and variations in TF residence time had a stronger influence than variations in concentration. This enabled us to device a model of gene transcription, in which TF binding triggers multiple successive steps before the gene transits to the active state and actual mRNA synthesis is decoupled from TF presence. We quantified all transition times of the TF and the gene, including the TF search time and the delay between TF binding and the onset of transcription. Our quantitative measurements and analysis revealed detailed kinetic insight, which may serve as basis for a bottom-up understanding of gene regulation.

scholarly journals Modulation of Nuclear Localization of the Influenza Virus Nucleoprotein through Interaction with Actin Filaments

1999 ◽  
Vol 73 (3) ◽  
pp. 2222-2231 ◽  
Author(s):  
Paul Digard ◽  
Debra Elton ◽  
Konrad Bishop ◽  
Elizabeth Medcalf ◽  
Alan Weeds ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Nuclear Localization ◽  
Virus Genome ◽  
Actin Binding ◽  
Filamentous Actin ◽  
Nuclear Localization Signals ◽  
Infected Cells ◽  
Cytoplasmic Accumulation ◽  
High Level

ABSTRACT The influenza virus genome is transcribed in the nuclei of infected cells but assembled into progeny virions in the cytoplasm. This is reflected in the cellular distribution of the virus nucleoprotein (NP), a protein which encapsidates genomic RNA to form ribonucleoprotein structures. At early times postinfection NP is found in the nucleus, but at later times it is found predominantly in the cytoplasm. NP contains several sequences proposed to act as nuclear localization signals (NLSs), and it is not clear how these are overridden to allow cytoplasmic accumulation of the protein. We find that NP binds tightly to filamentous actin in vitro and have identified a cluster of residues in NP essential for the interaction. Complexes containing RNA, NP, and actin could be formed, suggesting that viral ribonucleoproteins also bind actin. In cells, exogenously expressed NP when expressed at a high level partitioned to the cytoplasm, where it associated with F-actin stress fibers. In contrast, mutants unable to bind F-actin efficiently were imported into the nucleus even under conditions of high-level expression. Similarly, nuclear import of NLS-deficient NP molecules was restored by concomitant disruption of F-actin binding. We propose that the interaction of NP with F-actin causes the cytoplasmic retention of influenza virus ribonucleoproteins.

Mapping of the influenza virus genome II. Identification of the P1, P2, and P3 genes

Virology ◽  
1977 ◽  
Vol 76 (1) ◽  
pp. 114-121 ◽  
Author(s):  
Peter Palese ◽  
Mary B. Ritchey ◽  
Jerome L. Schulman
Keyword(s):  
Influenza Virus ◽  
Virus Genome

Gordon Leslie Ada 1922–2012

2015 ◽  
Vol 26 (2) ◽  
pp. 164
Author(s):  
Gustav Nossal ◽  
Christopher Parish
Keyword(s):  
Influenza Virus ◽  
Nobel Prize ◽  
Antibody Formation ◽  
Clonal Selection ◽  
Virus Genome ◽  
Research Environment ◽  
Clonal Selection Theory ◽  
Selection Theory ◽  
Viral Immunology ◽  
Administrative Skills

GordonAda, an outstanding virologist and immunologist,was the first to demonstrate that the influenza virus genome is composed of RNA, not DNA. In immunology he provided evidence that refuted the template theory of antibody formation and performed elegant experiments to prove Burnet's Clonal Selection Theory. His administrative skills created a research environment that nurtured the Doherty-Zinkernagel Nobel Prize in viral immunology and allowed him to assist WHO and others greatly in developing effective vaccines.

scholarly journals Kinetics of HTLV-1 reactivation from latency quantified by single-molecule RNA FISH and stochastic modelling

2019 ◽  
Vol 15 (11) ◽  
pp. e1008164 ◽  
Author(s):  
Michi Miura ◽  
Supravat Dey ◽  
Saumya Ramanayake ◽  
Abhyudai Singh ◽  
David S. Rueda ◽  
...  
Keyword(s):  
Single Molecule ◽  
Stochastic Modelling ◽  
Rna Fish ◽  
Kinetics Of

THE INFLUENZA VIRUS GENOME AND ITS REPLICATION

1979 ◽  
Vol 35 (1) ◽  
pp. 39-46 ◽  
Author(s):  
R D BARRY ◽  
B W J MAHY
Keyword(s):  
Influenza Virus ◽  
Virus Genome

scholarly journals Packaging of influenza virus genome: Robustness of selection

2012 ◽  
Vol 109 (23) ◽  
pp. 8797-8798 ◽  
Author(s):  
T. Noda ◽  
Y. Kawaoka
Keyword(s):  
Influenza Virus ◽  
Virus Genome
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