title: MiR-342-5p suppresses coxsackievirus B3 biosynthesis by targeting the 2C-coding region.
abstract: Coxsackievirus B type 3 (CVB3) is one of the major pathogens associated with human heart disease. miRNAs are a class of short, noncoding RNA that can post-transcriptionally modulate gene expression. By comparing the CVB3 genome and miR-342-5p sequences, we found there were potential miR-342-5p targets in the CVB3 genome. To verify the effect of miR-342-5p on CVB3 biosynthesis, HeLa cells were infected with a Renilla luciferase (RLuc)-expressing CVB3 variant (RLuc-CVB3). We observed that miR-342-5p could significantly inhibit the expression of RLuc in infected cells. In HeLa cells infected with an enhanced green fluorescence protein (EGFP)-expressing CVB3 variant (EGFP-CVB3), EGFP expression was also significantly inhibited by miR-342-5p. The inhibitory effect of miR-342-5p on EGFP expression in EGFP-CVB3-infected cells could be reversed by transfection with anti-miR-342-5p oligonucleotide (AMO-miR-342-5p). Moreover, RNA and protein biosynthesis in wild-type CVB3 was significantly inhibited by miR-342-5p. By mutating the putative targets of miR-342-5p in the 2C-coding region, a sequence, nt4989-nt5015, was identified as the miR-342-5p target. The conserved nt4989-nt5015 sequences of CVB type 1-5 suggest miR-342-5p may exert its inhibitory effect in other types of coxsackievirus besides CVB3. Western blotting indicated that miR-342-5p could indeed suppress protein expression in CVB type 1 and 5. There was a moderate abundance of miR-342-5p in the gut, heart, and brain of Balb/c mice, suggesting that miR-342-5p may interact with CVB3 in vivo. Taken together, these results indicate that miR-342-5p can inhibit CVB3 biosynthesis by targeting its 2C-coding region and therefore may be a potential therapeutic agent in the treatment of CVB3 infection.
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China; Department of Laboratory Diagnosis, The First Hospital of Harbin Medical University, Harbin 150001, China.
authors.forename: Linlin
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Tong
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Wu
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Qingguo
authors.initials: Q
authors.lastname: Jiao
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Lexun
authors.initials: L
authors.lastname: Lin
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China.
authors.forename: Ruixue
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Cell Biology, Harbin Medical University, Harbin 150081, China. Electronic address: wenran.zhao@gmail.com.
authors.forename: Wenran
authors.initials: W
authors.lastname: Zhao
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin 150081, China. Electronic address: zhongzh@hrbmu.edu.cn.
authors.forename: Zhaohua
authors.initials: Z
authors.lastname: Zhong
mesh_terms: D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D003384:Coxsackievirus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D015536:Down-Regulation; D029822:Enterovirus B, Human / Q000235:genetics* / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D014176:Protein Biosynthesis; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; C546176:MIRN342 microRNA, human; D035683:MicroRNAs; D017361:Viral Nonstructural Proteins; C076023:2C protein, viral