A1 Journal article (refereed)
Real-time Fluorescence Measurement of Enterovirus Uncoating (2020)
Ruokolainen, V., Laajala, M., & Marjomäki, V. (2020). Real-time Fluorescence Measurement of Enterovirus Uncoating. Bio-protocol, 10(7), Article e3582. https://doi.org/10.21769/BioProtoc.3582
JYU authors or editors
Publication details
All authors or editors: Ruokolainen, Visa; Laajala, Mira; Marjomäki, Varpu
Journal or series: Bio-protocol
eISSN: 2331-8325
Publication year: 2020
Volume: 10
Issue number: 7
Article number: e3582
Publisher: Bio-protocol
Publication country: United States
Publication language: English
DOI: https://doi.org/10.21769/BioProtoc.3582
Publication open access: Not open
Publication channel open access:
Publication is parallel published (JYX): https://jyx.jyu.fi/handle/123456789/68728
Additional information: The authors used this protocol in:
Ruokolainen, V., Domanska, A., Laajala, M., Pelliccia, M., Butcher, S. J., & Marjomäki, V. (2019). Extracellular Albumin and Endosomal Ions Prime Enterovirus Particles for Uncoating That Can Be Prevented by Fatty Acid Saturation. Journal of virology, 93(17), e00599-19. https://doi.org/10.1128/JVI.00599-19
Abstract
and translation. Especially for non-enveloped viruses, such as enteroviruses, the cues leading to
uncoating are less well known. The status of the virus has previously been observed mainly by
transmission electron microscopy using negative staining, cryo electron microscopy, X-ray
crystallography or gradient separation (reviewed in Tuthill et al., 2010, Myllynen et al., 2016,
Ruokolainen et al., 2019). However, monitoring of uncoating has been limited by the lack of methods
detecting dynamic changes of the virions. Here, we present a real-time fluorescence based protocol,
which detects the viral genome (RNA) during various stages of uncoating in vitro, while RNA is still inside
the particle that has been expanded before the actual RNA release, and when the RNA has been totally
released from the viral particle. Our method allows to explore how various molecular factors may
promote or inhibit virus uncoating.
Keywords: enteroviruses; infections; RNA; spectroscopy; fluorescence; research methods
Free keywords: Picornavirus; Enterovirus; uncoating; genome; RNA; RNase; SYBR Green II; fluorescence spectroscopy
Contributing organizations
Ministry reporting: Yes
Reporting Year: 2020
JUFO rating: 0