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Roles of bta-miR-29b promoter regions DNA methylation in regulating miR-29b expression and bovine viral diarrhea virus NADL replication in MDBK cells

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Abstract

MicroRNAs (miRNAs) are an important class of small, non-coding RNAs that control target genes expression by degradation of target mRNAs or by inhibiting protein translation in many biological processes and cellular pathways. In a previous study, we found that miR-29b interfered with bovine viral diarrhea virus (BVDV) replication. However, the mechanisms of regulation of miR-29b expression are not well known. DNA methylation is an important epigenetic mechanism for silencing gene transcription, and plays an important role in promoter choice, protein expression, and regulation of miRNAs expression. In this study, we focused on the roles of DNA methylation of miR-29b promoter in regulating miR-29b expression and investigated the effects of DNA (cytosine-5) methyltransferase 1 (DNMT1) knockdown on miR-29b expression and BVDV (strain NADL) replication. Our results showed that methylation levels of miR-29b promoter were significantly decreased in BVDV NADL-infected MDBK cells. Furthermore, DNMT1 silencing significantly decreased the methylation levels of miR-29b promoter, up-regulated miR-29b expression and inhibited BVDV NADL replication, which supports the important roles of DNA methylation in regulating miRNA expression and further proves an evidence for our previous views.

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Acknowledgments

This work was supported by the Natural Science Foundation of China (Grant Nos. 31502095, 31560328 and U1303283) and the International S&T Cooperation Program of China (Grant No. 2013DFR30970).

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Correspondence to Huijun Shi or Chuangfu Chen.

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Supplementary material 1 (DOCX 219 kb) Supplementary figure DNMT1 expression in BVDV NADL infected MDBK cells is significantly reduced compared with negative control. mRNA levels of DNMT1 were evaluated using RT-PCR in normal (negative control) and BVDV NADL-infected (100 TCID50/0.1 mL) MDBK cells for the indicated time intervals. The results are shown as the mean ± SD (error bars). *, P < 0.05, **, P < 0.01; n = 3

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Fu, Q., Shi, H. & Chen, C. Roles of bta-miR-29b promoter regions DNA methylation in regulating miR-29b expression and bovine viral diarrhea virus NADL replication in MDBK cells. Arch Virol 162, 401–408 (2017). https://doi.org/10.1007/s00705-016-3107-1

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