ID 57482
FullText URL
Author
Miyoshi, Yuichi Graduate School of Natural Science and Technology, Okayama University
Ohtsuki, Takashi Graduate School of Natural Science and Technology, Okayama University
Kashida, Hiromu Graduate School of Engineering, Nagoya University
Asanuma, Hiroyuki Graduate School of Engineering, Nagoya University
Watanabe, Kazunori Graduate School of Natural Science and Technology, Okayama University ORCID Kaken ID publons researchmap
Abstract
Cellular functions are regulated by the up- and down-regulation and localization of RNA molecules. Therefore, many RNA detection methods have been developed to analyze RNA levels and localization. Molecular beacon (MB) is one of the major methods for quantitative RNA detection and analysis of RNA localization. Most oligonucleotide-based probes, including MB, are designed to target a long flexible region on the target RNA molecule, e.g., a single-stranded region. Recently, analyses of tRNA localization and levels became important, as it has been shown that environmental stresses and chemical reagents induce nuclear accumulation of tRNA and tRNA degradation in mammalian cells. However, tRNA is highly structured and does not harbor any long flexible regions. Hence, only a few methods are currently available for detecting tRNA. In the present study, we attempted to detect elongator tRNAMet (eMet) and initiator tRNAMet (iMet) by using an in-stem molecular beacon (ISMB), characterized by more effective quenching and significantly higher sensitivity than those of conventional MB. We found that ISMB1 targeted a 5'- region that includes the D arm of tRNA and that it detected eMet and iMet transcripts as well as mature eMet with high sensitivity. Moreover, the analysis revealed that the formation of the ISMB/tRNA transcript complex required more time than the formation of an ISMB/unstructured short RNA complex. These results suggest that ISMB-based tRNA detection can be a useful tool for various biological and medical studies.
Published Date
2019-01-29
Publication Title
PLoS One
Volume
volume14
Issue
issue1
Publisher
PUBLIC LIBRARY SCIENCE
Start Page
e0211505
ISSN
1932-6203
Content Type
Journal Article
language
フランス語
OAI-PMH Set
岡山大学
Copyright Holders
© 2019 Miyoshi et al.
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PubMed ID
DOI
Web of Science KeyUT
Related Url
isVersionOf https://doi.org/10.1371/journal.pone.0211505
License
https://creativecommons.org/licenses/by/4.0/
Citation
Miyoshi Y, Ohtsuki T, Kashida H, Asanuma H, Watanabe K (2019) In-stem molecular beacon targeted to a 50 -region of tRNA inclusive of the D arm that detects mature tRNA with high sensitivity. PLoS ONE 14(1): e0211505. https://doi. org/10.1371/journal.pone.0211505
Funder Name
Japan Society for the Promotion of Science
助成番号
26840010
16H05925
17H05150
18H03933
15K13746