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Discoveries of Exoribonuclease-Resistant Structures of Insect-Specific Flaviviruses Isolated in Zambia

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Title: Discoveries of Exoribonuclease-Resistant Structures of Insect-Specific Flaviviruses Isolated in Zambia
Authors: Wastika, Christida E. Browse this author
Harima, Hayato Browse this author
Sasaki, Michihito Browse this author
Hang'ombe, Bernard M. Browse this author
Eshita, Yuki Browse this author
Qiu, Yongjin Browse this author
Hall, William W. Browse this author
Wolfinger, Michael T. Browse this author
Sawa, Hirofumi Browse this author
Orba, Yasuko Browse this author →KAKEN DB
Keywords: Barkedji virus
Barkedji-like virus
5 '-UTR
3 '-UTR
RNA secondary structure
Issue Date: Sep-2020
Publisher: MDPI
Journal Title: Viruses-Basel
Volume: 12
Issue: 9
Start Page: 1017
Publisher DOI: 10.3390/v12091017
Abstract: To monitor the arthropod-borne virus transmission in mosquitoes, we have attempted both to detect and isolate viruses from 3304 wild-caught female mosquitoes in the Livingstone (Southern Province) and Mongu (Western Province) regions in Zambia in 2017. A pan-flavivirus RT-PCR assay was performed to identify flavivirus genomes in total RNA extracted from mosquito lysates, followed by virus isolation and full genome sequence analysis using next-generation sequencing and rapid amplification of cDNA ends. We isolated a newly identified Barkedji virus (BJV Zambia) (10,899 nt) and a novel flavivirus, tentatively termed Barkedji-like virus (BJLV) (10,885 nt) fromCulexspp. mosquitoes which shared 96% and 75% nucleotide identity with BJV which has been isolated in Israel, respectively. These viruses could replicate in C6/36 cells but not in mammalian and avian cell lines. In parallel, a comparative genomics screening was conducted to study evolutionary traits of the 5 '- and 3 '-untranslated regions (UTRs) of isolated viruses. Bioinformatic analyses of the secondary structures in the UTRs of both viruses revealed that the 5 '-UTRs exhibit canonical stem-loop structures, while the 3 '-UTRs contain structural homologs to exoribonuclease-resistant RNAs (xrRNAs), SL-III, dumbbell, and terminal stem-loop (3 ' SL) structures. The function of predicted xrRNA structures to stop RNA degradation by Xrn1 exoribonuclease was further proved by the in vitro Xrn1 resistance assay.
Type: article
Appears in Collections:人獣共通感染症リサーチセンター (Research Center for Zoonosis Control) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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