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MicroRNA miR-320a and miR-140 inhibit mink enteritis virus infection by repression of its receptor, feline transferrin receptor.

Sun JZ, Wang J, Wang S, Yuan D, Li Z, Yi B, Hou Q, Mao Y, Liu W - Virol. J. (2014)

Bottom Line: Recent studies have shed light into the role of microRNAs (miRNAs), small noncoding RNAs of length ranging from 18-23 nucleotides (nt), as critical modulators in the host-pathogen interaction networks.We previously showed that miRNA miR-181b can inhibit MEV replication by repression of viral non-structural protein 1 expression.Here, we report that two other miRNAs (miR-320a and miR-140) inhibit MEV entry into feline kidney (F81) cells by downregulating its receptor, transferrin receptor (TfR), by targeting the 3' untranslated region (UTR) of TfR mRNA, while being themselves upregulated.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agrobiotechnology, Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100193, China. sunjiazeng331@163.com.

ABSTRACT
Mink enteritis virus (MEV) is one of the most important pathogens in the mink industry. Recent studies have shed light into the role of microRNAs (miRNAs), small noncoding RNAs of length ranging from 18-23 nucleotides (nt), as critical modulators in the host-pathogen interaction networks. We previously showed that miRNA miR-181b can inhibit MEV replication by repression of viral non-structural protein 1 expression. Here, we report that two other miRNAs (miR-320a and miR-140) inhibit MEV entry into feline kidney (F81) cells by downregulating its receptor, transferrin receptor (TfR), by targeting the 3' untranslated region (UTR) of TfR mRNA, while being themselves upregulated.

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MEV infection leads to gradual changes of miR-320a, miR-140 and TfR in F81 cells. qPCR was used to assess the relative expression of miR-320a, miR-140 and TfR mRNAs at the indicated times (A) under normal conditions or (B) after MEV infection, with β-actin as an internal control. Data are from 3 independent experiments (mean ± SD). (C) Western blot assay was used to assess the TfR expression levels under normal conditions (−) or after MEV infection (+). (D) Western blot assay was used to assess the TfR expression levels when transfection with miRNA inhibitors post 12 h MEV infection.
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Fig3: MEV infection leads to gradual changes of miR-320a, miR-140 and TfR in F81 cells. qPCR was used to assess the relative expression of miR-320a, miR-140 and TfR mRNAs at the indicated times (A) under normal conditions or (B) after MEV infection, with β-actin as an internal control. Data are from 3 independent experiments (mean ± SD). (C) Western blot assay was used to assess the TfR expression levels under normal conditions (−) or after MEV infection (+). (D) Western blot assay was used to assess the TfR expression levels when transfection with miRNA inhibitors post 12 h MEV infection.

Mentions: The expression levels of predicted miRNAs are shown in Table 1. Following MEV infection both miR-320a and miR-140 showed relatively higher level expression and greater upregulation than found in uninfected cells. This was confirmed by qPCR which showed that both miRNAs gradually increased following MEV infection (Figure 3). To investigate whether upregulation of the two miRNAs following MEV infection could affect TfR expression, qPCR (Figure 3A,B) and western blot (Figure 3C) were performed simultaneously. As expected, TfR was gradually downregulated following MEV infection. To further ascertain that TfR downregulation is the result of miRNAs upregulation, F81 cells were transfected with miRNA inhibitors 12 h post MEV infection. Results showed that compared with transfection with NC inhibitors, both miR-320a and miR-140 inhibitors attenuated the inhibitory effect on TfR expression levels (Figure 3D). It appears, therefore, that upregulation of miR-320a and miR-140 directly results in TfR downregulation.Table 1


MicroRNA miR-320a and miR-140 inhibit mink enteritis virus infection by repression of its receptor, feline transferrin receptor.

Sun JZ, Wang J, Wang S, Yuan D, Li Z, Yi B, Hou Q, Mao Y, Liu W - Virol. J. (2014)

MEV infection leads to gradual changes of miR-320a, miR-140 and TfR in F81 cells. qPCR was used to assess the relative expression of miR-320a, miR-140 and TfR mRNAs at the indicated times (A) under normal conditions or (B) after MEV infection, with β-actin as an internal control. Data are from 3 independent experiments (mean ± SD). (C) Western blot assay was used to assess the TfR expression levels under normal conditions (−) or after MEV infection (+). (D) Western blot assay was used to assess the TfR expression levels when transfection with miRNA inhibitors post 12 h MEV infection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4264318&req=5

Fig3: MEV infection leads to gradual changes of miR-320a, miR-140 and TfR in F81 cells. qPCR was used to assess the relative expression of miR-320a, miR-140 and TfR mRNAs at the indicated times (A) under normal conditions or (B) after MEV infection, with β-actin as an internal control. Data are from 3 independent experiments (mean ± SD). (C) Western blot assay was used to assess the TfR expression levels under normal conditions (−) or after MEV infection (+). (D) Western blot assay was used to assess the TfR expression levels when transfection with miRNA inhibitors post 12 h MEV infection.
Mentions: The expression levels of predicted miRNAs are shown in Table 1. Following MEV infection both miR-320a and miR-140 showed relatively higher level expression and greater upregulation than found in uninfected cells. This was confirmed by qPCR which showed that both miRNAs gradually increased following MEV infection (Figure 3). To investigate whether upregulation of the two miRNAs following MEV infection could affect TfR expression, qPCR (Figure 3A,B) and western blot (Figure 3C) were performed simultaneously. As expected, TfR was gradually downregulated following MEV infection. To further ascertain that TfR downregulation is the result of miRNAs upregulation, F81 cells were transfected with miRNA inhibitors 12 h post MEV infection. Results showed that compared with transfection with NC inhibitors, both miR-320a and miR-140 inhibitors attenuated the inhibitory effect on TfR expression levels (Figure 3D). It appears, therefore, that upregulation of miR-320a and miR-140 directly results in TfR downregulation.Table 1

Bottom Line: Recent studies have shed light into the role of microRNAs (miRNAs), small noncoding RNAs of length ranging from 18-23 nucleotides (nt), as critical modulators in the host-pathogen interaction networks.We previously showed that miRNA miR-181b can inhibit MEV replication by repression of viral non-structural protein 1 expression.Here, we report that two other miRNAs (miR-320a and miR-140) inhibit MEV entry into feline kidney (F81) cells by downregulating its receptor, transferrin receptor (TfR), by targeting the 3' untranslated region (UTR) of TfR mRNA, while being themselves upregulated.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agrobiotechnology, Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100193, China. sunjiazeng331@163.com.

ABSTRACT
Mink enteritis virus (MEV) is one of the most important pathogens in the mink industry. Recent studies have shed light into the role of microRNAs (miRNAs), small noncoding RNAs of length ranging from 18-23 nucleotides (nt), as critical modulators in the host-pathogen interaction networks. We previously showed that miRNA miR-181b can inhibit MEV replication by repression of viral non-structural protein 1 expression. Here, we report that two other miRNAs (miR-320a and miR-140) inhibit MEV entry into feline kidney (F81) cells by downregulating its receptor, transferrin receptor (TfR), by targeting the 3' untranslated region (UTR) of TfR mRNA, while being themselves upregulated.

Show MeSH
Related in: MedlinePlus