A1 Journal article (refereed)
Real-time Fluorescence Measurement of Enterovirus Uncoating (2020)


Ruokolainen, Visa; Laajala, Mira; Marjomäki, Varpu (2020). Real-time Fluorescence Measurement of Enterovirus Uncoating. Bio-protocol, 10 (7), e3582. DOI: 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

Open Access: Publication channel is not openly available

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

Viruses need to open, i.e., uncoat, in order to release their genomes for efficient replication
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

Preliminary JUFO rating: 0


Last updated on 2020-18-08 at 13:43