Over expression of bmo-miR-2819 suppresses BmNPV replication by regulating the BmNPV ie-1 gene in Bombyx mori

Wu, Ping; Shang, Qi; Dweteh, Owusu Amanfo; Huang, Haoling; Zhang, Shaolun; Zhong, Jinbo; Hou, Qirui; Guo, Xingchen

doi:10.1016/j.molimm.2019.03.013

“…In order to detect the relative expression of host genes in different treatments, relative RT-qPCR was performed as described in one of our previous studies (Wu et al, 2019b). In order to determine the BmNPV gene and vDNA synthesis amounts, absolute RT-qPCR was carried out as described in another of our previous studies (Wu et al, 2019a). Three independent experiments with three technical replicates were performed.…”

Section: Rt-qpcr Analysismentioning

confidence: 99%

Inhibition of heat shock protein 90 suppresses Bombyx mori nucleopolyhedrovirus replication in B. mori

Shang

¹

,

Wu

²

,

Huang

³

et al. 2019

Insect Molecular Biology

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Heat shock protein 90 (Hsp90) plays a very important role in facilitating the replication of many viruses. Until now, little has been known about the role of Hsp90 in Bombyx mori virus infection. In this study, we explored the role of BmHsp90 in B. mori nucleopolyhedrovirus (BmNPV) replication. We found that BmHsp90 inhibition by geldanamycin (GA) significantly reduced the BmNPV titre, the protein expression level of BmNPV nucleocapsid protein 39 (VP39) and the transcript level of BmNPV genes. Silencing the hsp90 gene in BmN cells by small interfering RNA suppressed BmNPV replication whereas overexpression of hsp90 promoted the replication of BmNPV. After inhibition of Hsp90, the expression of three key genes [signal transducing activator of transcription (stat), suppressor of cytokine signalling protein 2 (socs2), socs6] involved in the Janus kinase/STAT pathway significantly changed, with up‐regulation of stat and down‐regulation of socs2 and socs6. In addition, the expression of two antiapoptosis genes, BmNPV inhibitor of apoptosis protein1 (BmNPV‐iap1) and Bmiap2, was greatly decreased in GA‐treated cells, whereas their expression was significantly increased in hsp90‐overexpressed silkworm larvae. Our results indicated that inhibition of Hsp90 can suppress BmNPV proliferation in B. mori. Our findings may provide new clues to elucidate the molecular mechanisms of silkworm–virus interactions.

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“…Relative quantitative real-time PCR (qRT-PCR) as descried in our previous study was performed to detect the expression of silkworm gene [63,64]. As to BmNPV gene lef-3 (GeneID: 1488686) and ie-1 (GeneID: 1488755), absolute qRT-PCR was carried out as in our previous report [63]. PCR reactions were run with a thermal cycling at 95 °C for 30 s followed by 40 cycles of 95 °C for 5 s, 60 °C for 31 s.…”

Section: Quantitative Real-time Pcr Analysismentioning

confidence: 99%

“…Western blotting assay was performed as descried in our previous study [63]. Briefly, a total of 30 µg protein sample from cells was loaded on each lane and separated on SDS-PAGE before transferred onto a nitrocellulose membrane.…”

Section: Western Blot Assaymentioning

confidence: 99%

DNA methylomes and transcriptomes analysis reveal implication of host DNA methylation machinery in BmNPV proliferation in Bombyx mori

Huang

¹

,

Wu

²

,

Zhang

³

et al. 2019

Preprint

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Background Bombyx mori nucleopolyhedrosis virus (BmNPV) is a major pathogen that threatens the sustainability of the sericultural industry. DNA methylation is a widespread gene regulation mode in epigenetics, which plays an important role in host immune response. Until now, little has been known about epigenetic regulation on virus diseases in insects. This study aims to explore the role of DNA methylation in BmNPV proliferation.Results Inhibiting DNA methyltransferase (DNMT) activity of silkworm can suppress BmNPV replication. The integrated analysis of transcriptomes and DNA methylomes in silkworm midguts infected with or without BmNPV showed that both the expression pattern of transcriptome and DNA methylation pattern are changed significantly upon BmNPV infection. A total of 291 differentially methylated genes (DMGs) were observed in BmNPV infected midguts, among which, 126 genes were hypermethylated and 115 genes were hypomethylated. Significant differences in both mRNA transcript level and DNA methylated levels were found in 26 genes. BS-PCR validated the hypermethylation of BGIBMGA014008 , a structural maintenance of chromosomes protein gene in the BmNPV-infected midgut. In addition, DNMT inhibition reduced the expression of inhibitor of apoptosis family genes, iap1 from BmNPV, Bmiap2, BmSurvivin1 and BmSurvivin2 .Conclusion Our results indicate that DNA methylation plays positive roles in BmNPV proliferation and loss of DNMT activity could induce the apoptosis of infected cells to suppress BmNPV proliferation. Our results may provide a new idea and research direction for the molecular mechanism on insect-virus interaction.

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“…Relative quantitative real-time PCR (qRT-PCR) as descried in our previous study was performed to detect the expression of silkworm gene [64,65]. As to BmNPV gene lef-3 (GeneID: 1488686) and ie-1 (GeneID: 1488755), absolute qRT-PCR was carried out as in our previous report [64]. PCR reactions were run with a thermal cycling at 95 °C for 30 s followed by 40 cycles of 95 °C for 5 s, 60 °C for 31 s.…”

Section: Quantitative Real-time Pcr Analysismentioning

confidence: 99%

DNA methylomes and transcriptomes analysis reveal implication of host DNA methylation machinery in BmNPV proliferation in Bombyx mori

Huang

¹

,

Wu

²

,

Zhang

³

et al. 2019

Preprint

Self Cite

0

View full text Add to dashboard Cite

Background Bombyx mori nucleopolyhedrosis virus (BmNPV) is a major pathogen that threatens the sustainability of the sericultural industry. DNA methylation is a widespread gene regulation mode in epigenetics, which plays an important role in host immune response. Until now, little has been known about epigenetic regulation on virus diseases in insects. This study aims to explore the role of DNA methylation in BmNPV proliferation.Results Inhibiting DNA methyltransferase (DNMT) activity of silkworm can suppress BmNPV replication. The integrated analysis of transcriptomes and DNA methylomes in silkworm midguts infected with or without BmNPV showed that both the expression pattern of transcriptome and DNA methylation pattern are changed significantly upon BmNPV infection. A total of 291 differentially methylated genes (DMGs) were observed in BmNPV infected midguts, among which, 126 genes were hypermethylated and 115 genes were hypomethylated. Significant differences in both mRNA transcript level and DNA methylated levels were found in 26 genes. BS-PCR validated the hypermethylation of BGIBMGA014008 , a structural maintenance of chromosomes protein gene in the BmNPV-infected midgut. In addition, DNMT inhibition reduced the expression of inhibitor of apoptosis family genes, iap1 from BmNPV, Bmiap2, BmSurvivin1 and BmSurvivin2 .Conclusion Our results indicate that DNA methylation plays positive roles in BmNPV proliferation and loss of DNMT activity could induce the apoptosis of infected cells to suppress BmNPV proliferation. Our results may provide a new idea and research direction for the molecular mechanism on insect-virus interaction.

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Over expression of bmo-miR-2819 suppresses BmNPV replication by regulating the BmNPV ie-1 gene in Bombyx mori

Cited by 36 publications

References 28 publications

Inhibition of heat shock protein 90 suppresses Bombyx mori nucleopolyhedrovirus replication in B. mori

Inhibition of heat shock protein 90 suppresses Bombyx mori nucleopolyhedrovirus replication in B. mori

DNA methylomes and transcriptomes analysis reveal implication of host DNA methylation machinery in BmNPV proliferation in Bombyx mori

DNA methylomes and transcriptomes analysis reveal implication of host DNA methylation machinery in BmNPV proliferation in Bombyx mori

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