Characterization of cDNAs encoding p53 of Bombyx mori and Spodoptera frugiperda

Huang, Ning; Clem, Rollie J.; Rohrmann, George F.

doi:10.1016/j.ibmb.2011.03.014

Cited by 21 publications

(30 citation statements)

References 35 publications

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“…They were then washed and incubated with horseradish peroxidase-conjugated secondary antibodies (Sigma). Signal development was conducted as described previously (16). All immunoblotting experiments were done multiple times using samples from at least three biological replicates, and representative results are presented.…”

Section: Cells and Virusesmentioning

confidence: 99%

“…Caspase activity was determined using the mammalian caspase-3 substrate N-acetyl-Asp-Glu-Val-Asp-(7-amino-4-trifluoromethylcoumarin) (DEVD-AFC; MP Biomedicals) as described previously (16). Cell viability was determined by photographing four random fields of view at ϫ400 magnification at the indicated times and counting the number of cells with normal (nonapoptotic) morphology relative to untreated cells (set at 100%).…”

Section: Cells and Virusesmentioning

confidence: 99%

“…Proteins were transferred to a PVDF membrane using CAPS buffer (10 mM CAPS [Sigma], pH 11, 20% methanol). PVDF membranes containing transferred protein were blocked with 5% bovine serum albumin for 1 h, incubated with anti-phospho-histone H2AX polyclonal antibody (catalog number PA-40278; Pierce) diluted 1:5,000 overnight at 4°C, washed, and incubated with peroxidase-labeled goat anti-rabbit secondary antibody (Abcam) at a 1:5,000-fold dilution for 1.5 h. Signal development was performed as described previously (16).…”

Section: Cells and Virusesmentioning

confidence: 99%

“…SfP53 shares low but significant sequence similarity with DmP53, including conserved amino acids involved in DNA binding. Overexpression of SfP53 was shown to stimulate apoptosis, but unlike DmP53, levels of endogenous SfP53 protein were increased when cells of Sf9, a S. frugiperda cell line, were treated with the DNA-damaging agents UV or camptothecin (CPT) (16). This increased protein abundance is similar to what is seen for P53 in mammalian cells following DNA damage, but whether it is due to increased transcription or altered protein stability is not known.…”

mentioning

confidence: 95%

“…A lepidopteran ortholog of p53, Sfp53, was recently identified in the fall armyworm Spodoptera frugiperda, a host of AcMNPV (16). SfP53 shares low but significant sequence similarity with DmP53, including conserved amino acids involved in DNA binding.…”

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confidence: 99%

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Baculovirus Infection Induces a DNA Damage Response That Is Required for Efficient Viral Replication

Huang

Yang

et al. 2011

J Virol

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Several mammalian viruses have been shown to induce a cellular DNA damage response during replication, and in some cases, this response is required for optimal virus replication. However, nothing is known about whether a DNA damage response is stimulated by DNA viruses in invertebrates. Cell cycle arrest and apoptosis are two of the downstream effects of the DNA damage response, and both are stimulated by baculovirus infection, suggesting a possible relationship between baculoviruses and the DNA damage response. In the study described in this report, we found that replication of the baculovirus Autographa californica M nucleopolyhedrovirus (AcMNPV) in the cell line Sf9, derived from the lepidopteran insect Spodoptera frugiperda, stimulated a DNA damage response, as indicated by an increased abundance of the S. frugiperda P53 protein (SfP53) and phosphorylation of the histone variant protein H2AX. Stimulation of the DNA damage response was dependent on viral DNA replication. Inhibition of the DNA damage response prevented both the increase in SfP53 accumulation and H2AX phosphorylation and also caused a 10-to 100-fold reduction in virus production, along with decreased viral DNA replication and late gene expression. However, silencing of Sfp53 expression by RNA interference did not significantly affect AcMNPV replication or induction of apoptosis by a mutant of AcMNPV lacking the antiapoptotic gene p35, indicating that these processes are not dependent on SfP53 in Sf9 cells.Eukaryotic cells constantly monitor the integrity of their genomes and rapidly respond to the presence of damaged DNA by activating DNA damage response pathways, which are initiated largely through the activation of two members of the phosphatidylinositol 3-kinase superfamily, ataxia telangiectasia mutated (ATM) and ATM and rad-3 related (ATR) (for a recent review, see reference 50). ATM is activated mainly in response to double-strand breaks in DNA, while ATR responds mainly to single-strand breaks and stalled replication forks. However, there is overlap in the downstream substrates of these kinases. ATM and ATR phosphorylate numerous effector proteins which function in cell cycle checkpoints, DNA repair, and stimulation of apoptosis. One of the many important substrates of ATM and ATR is the transcription factor P53, which regulates the expression of numerous genes that function in all three of these processes. Studies in Drosophila melanogaster have demonstrated that DNA damage responses, including the roles of ATM, ATR, and P53, share many similarities in insects and mammals (53).There is increasing evidence that the ability to manipulate the DNA damage response and associated downstream pathways is crucial for the replication of many viruses. Several mammalian viruses stimulate DNA damage response pathways as a consequence of infection, and these viruses have in turn evolved mechanisms to manipulate these pathways for their own benefit by exploiting or actively inhibiting different parts of the pathways (reviewed in reference 7). For exam...

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Section: Cells and Virusesmentioning

confidence: 99%

Section: Cells and Virusesmentioning

confidence: 99%

Section: Cells and Virusesmentioning

confidence: 99%

mentioning

confidence: 95%

mentioning

confidence: 99%

See 3 more Smart Citations

Baculovirus Infection Induces a DNA Damage Response That Is Required for Efficient Viral Replication

Huang

Yang

et al. 2011

J Virol

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Baculovirus antiapoptotic protein P35 regulated the host apoptosis to enhance virus multiplication

Miao

Liang

2016

Mol Cell Biochem

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The baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) possesses p35 gene, which encodes antiapoptosis protein P35 (or Ac-P35). A previous study showed that deleting p35 promoted premature cytolysis and caused the reduction of virus yield. In this report, we examined the role of P35 in regulating the expression of Ac-IAPs (inhibitor of apoptosis proteins in AcMNPV) and SfP53 (an apoptosis protein in Sf9 cells), and its effect on the production of progeny virus. Results showed that the overexpression of P35 caused a delay in the increase process of SfP53 before 36-h post infection and improved the transcription levels of iaps gene dramatically; it was more favorable to improve the transcription level of iap1 at 24-72 h post infection. Moreover, P35 could promote the production of progeny virus. This is the first study to disclose the relationship among p35, iap1, iap2, Sfp53 and the replication of the virus in the AcMNPV infection process, which provides the basis for improving the insecticidal activity of recombinant AcMNPV in terms of theory and technology.

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Gene expression pattern of insect fat body cells from in vitro challenge to cell line establishment

Zhang

Meng

Tang

et al. 2014

In Vitro Cell.Dev.Biol.-Animal

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The cell lines provided excellent tools to understand the mechanism of biological phenomenon at the cellular and molecular levels. The continuous development of new cell culture technology is both of interest for use in biochemical, immunology, and virological studies. The transformation of cells of the primary culture is a key procedure for insect cell line establishment but little is known about the molecular basis of these changes. Here, we found that the cell cycle progression of the cells of the primary culture was delayed or arrested in G2/M by fluorescence-activated cell sorting analysis. In this study, two subtractive cDNA libraries were constructed to screen for immortal-related genes of Spodoptera exigua (Lepidoptera: Noctuidae). Gene ontology and pathway analysis indicated that members of the oxidative phosphorylation, PI3K-Akt signaling pathway, and the ubiquitin proteasome pathway are involved in processes leading toward cell immortalization merit further investigation. Our findings suggest that tumor-related genes or target genes of these pathways may contribute to the transformation of primary cell through regulation of G2/M cell cycle progression.

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Characterization of cDNAs encoding p53 of Bombyx mori and Spodoptera frugiperda

Cited by 21 publications

References 35 publications

Baculovirus Infection Induces a DNA Damage Response That Is Required for Efficient Viral Replication

Baculovirus Infection Induces a DNA Damage Response That Is Required for Efficient Viral Replication

Baculovirus antiapoptotic protein P35 regulated the host apoptosis to enhance virus multiplication

Gene expression pattern of insect fat body cells from in vitro challenge to cell line establishment

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