Novel Macula-Like Virus Identified in
            <i>Bombyx mori</i>
            Cultured Cells

Katsuma, Susumu; Tanaka, Shinichiro; Omuro, Naoko; Takabuchi, Lisa; Daimon, Takaaki; Imanishi, Shigeo; Yamashita, Shizuya; Iwanaga, Masako; Mita, Kazuei; Maeda, Susumu; Kobayashi, Masahiko; Shimada, Toru

doi:10.1128/jvi.79.9.5577-5584.2005

Cited by 71 publications

(88 citation statements)

References 16 publications

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“…mori larvae (F1 hybrid Kinshu × Showa) were reared as described previously (Katsuma et al, 2005). The Sf-9 and High Five cell lines were cultured at 27°C in TC-100 medium supplemented with 10% fetal bovine serum and Express Five medium (Invitrogen, Carlsbad, CA), respectively.…”

Section: Insects Cell Lines and Virusesmentioning

confidence: 99%

Functional analysis of four Gloverin‐like genes in the silkworm, Bombyx mori

Kawaoka

Katsuma

Daimon

et al. 2007

Arch Insect Biochem Physiol

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To identify genes involved in the innate immunity of the silkworm Bombyx mori, we constructed a cDNA library from the fat body of Escherichia coli-challenged B. mori larvae. Based on the expressed sequence tag (EST) data and whole genome shotgun sequence analysis, we found four Gloverin-like genes, BmGlov1-4, in the Bombyx genome. Northern blot and RT-PCR analysis showed that BmGlov1-4 were induced in the larval fat body after an immune challenge by the injection of E. coli; however, less induction was observed after the injection of a yeast Candida albicans. In silico sequence analysis revealed the presence of a motif homologous to NF-kappaB binding site in the upstream region of each BmGlov gene. Moreover, we expressed recombinant BmGlov1-4 proteins using the baculovirus expression system, and found that all the recombinant BmGlov1-4 significantly inhibited the growth of E. coli.

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Section: Insects Cell Lines and Virusesmentioning

confidence: 99%

Functional analysis of four Gloverin‐like genes in the silkworm, Bombyx mori

Kawaoka

Katsuma

Daimon

et al. 2007

Arch Insect Biochem Physiol

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“…Two days after transfection, zeocin (final concentration, 500 mg/mL) was added to the medium. Two weeks after drug selection, the expression of FLAG-tagged Siwi and BmAgo3 was verified using the previously described Western blot analysis (Katsuma et al 2005) with anti-FLAG monoclonal (1:1000, Sigma), anti-Siwi (1:10,000), or antiBmAgo3 (1:5000) antibodies.…”

Section: Generation Of Bmn4 Cells Stably Expressing Flag/his-tagged Pmentioning

confidence: 99%

The Bombyx ovary-derived cell line endogenously expresses PIWI/PIWI-interacting RNA complexes

Kawaoka¹,

Hayashi²,

Suzuki³

et al. 2009

RNA

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Genetic studies and large-scale sequencing experiments have revealed that the PIWI subfamily proteins and PIWI-interacting RNAs (piRNAs) play an important role in germ line development and transposon control. Biochemical studies in vitro have greatly contributed to the understanding of small interfering RNA (siRNA) and microRNA (miRNA) pathways. However, in vitro analyses of the piRNA pathway have been thus far quite challenging, because their expression is largely restricted to the germ line. Here we report that Bombyx mori ovary-derived cultured cell line, BmN4, endogenously expresses two PIWI subfamily proteins, silkworm Piwi (Siwi) and Ago3 (BmAgo3), and piRNAs associated with them. Siwi-bound piRNAs have a strong bias for uridine at their 59 end and BmAgo3-bound piRNAs are enriched for adenine at position 10. In addition, Siwi preferentially binds antisense piRNAs, whereas BmAgo3 binds sense piRNAs. Moreover, we identified many pairs in which Siwi-bound antisense and BmAgo3-bound sense piRNAs are overlapped by precisely 10 nt at their 59 ends. These signatures are known to be important for secondary piRNA biogenesis in other organisms. Taken together, BmN4 is a unique cell line in which both primary and secondary steps of piRNA biogenesis pathways are active. This cell line would provide useful tools for analysis of piRNA biogenesis and function.

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“…Infection typically causes a global shutoff of host protein synthesis and gene expression in insect cells beginning at around 12 to 18 h postinfection (p.i.) (hpi) (29,42,46). Conversely, particular host genes are induced or remain stably expressed until the late stage of baculovirus infection.…”

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confidence: 99%

ERK- and JNK-Dependent Signaling Pathways Contribute to Bombyx mori Nucleopolyhedrovirus Infection

Katsuma

Mita

Shimada

2007

J Virol

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110

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Mitogen-activated protein kinases (MAPKs) often play important roles in virus infection. To explore intracellular signaling pathways induced by baculovirus infection, we examined the involvement of MAPKs inBombyx mori nucleopolyhedrovirus (BmNPV) infection of BmN cells. We found that specific inhibitors of extracellular signal-regulated kinase (ERK) kinase and c-Jun NH 2 -terminal kinase (JNK) significantly reduced occlusion body (OB) formation and budded virus (BV) production. Next, we quantified OB and BV production after applying the inhibitors at different times postinfection (p.i.). The inhibitors significantly reduced OB and BV production to various extents when applied at 12 h p.i., indicating that the reduction of BmNPV infectivity by these inhibitors occurs at the late stage of infection. Also, we observed that these inhibitors markedly repressed or deregulated the expression of delayed early, late, and very late gene products. Western blot analysis using phospho-MAPK-specific antibodies showed that ERK and JNK were activated at the late stage of BmNPV infection. In addition, the magnitude and pattern of MAPK activation were dependent on the multiplicity of infection. To verify the effects of the inhibitors on BmNPV infection, we also attempted to knock down the B. mori genes BmErk and BmJnk, which encode ERK and JNK, respectively. Knockdown of BmErk and BmJnk resulted in the reduced production of OBs and BVs, confirming that BmERK and BmJNK are involved in the BmNPV infection process. Taken together, these results indicate that the activation of MAPK signaling pathways is required for efficient infection by BmNPV.

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Novel Macula-Like Virus Identified in Bombyx mori Cultured Cells

Cited by 71 publications

References 16 publications

Functional analysis of four Gloverin‐like genes in the silkworm, Bombyx mori

Functional analysis of four Gloverin‐like genes in the silkworm, Bombyx mori

The Bombyx ovary-derived cell line endogenously expresses PIWI/PIWI-interacting RNA complexes

ERK- and JNK-Dependent Signaling Pathways Contribute to Bombyx mori Nucleopolyhedrovirus Infection

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