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Identification and characterization of the Spodoptera Su(var) 3-9 histone H3K9 trimethyltransferase and its effect in AcMNPV infection.

Li B, Li S, Yin J, Zhong J - PLoS ONE (2013)

Bottom Line: Su(var) 3-9 protein was found to be localized in the nucleus in Sf9 cells, and interact with histone H3, and the heterochromatin protein 1a (HP1a) and HP1b.A dose-dependent enzymatic activity was found at both 27 °C and 37 °C in vitro, with higher activity at 27 °C.Addition of specific inhibitor chaetocin resulted in decreased histone methylation level and host chromatin relaxation.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, People's Republic of China.

ABSTRACT
Histone H3-lysine(9) (H3K9) trimethyltransferase gene Su(var) 3-9 was cloned and identified in three Spodoptera insects, Spodopterafrugiperda (S. frugiperda), S. exigua and S. litura. Sequence analysis showed that Spodoptera Su(var) 3-9 is highly conserved evolutionarily. Su(var) 3-9 protein was found to be localized in the nucleus in Sf9 cells, and interact with histone H3, and the heterochromatin protein 1a (HP1a) and HP1b. A dose-dependent enzymatic activity was found at both 27 °C and 37 °C in vitro, with higher activity at 27 °C. Addition of specific inhibitor chaetocin resulted in decreased histone methylation level and host chromatin relaxation. In contrast, overexpression of Su(var) 3-9 caused increased histone methylation level and cellular genome compaction. In AcMNV-infected Sf9 cells, the transcription of Su(var) 3-9 increased at late time of infection, although the mRNA levels of most cellular genes decreased. Pre-treatment of Sf9 cells with chaetocin speeded up viral DNA replication, and increased the transcription level of a variety of virus genes, whereas in Sf9 cells pre-transformed with Su(var) 3-9 expression vector, viral DNA replication slow down slightly. These findings suggest that Su(var) 3-9 might participate in the viral genes expression an genome replication repression during AcMNPV infection. It provided a new insight for the understanding virus-host interaction mechanism.

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Related in: MedlinePlus

The effect of AcMNPV infection on host gene activity.Genomic DNA from AcMNPV or mock -infected cells were digested with DNase I. The percentage of DNA resistant to the nuclease was determined with qPCR with primers for S. frugiperda β-Actin and β-Tubulin. The error bars indicate the standard deviation calculated from at least three independent parallel experiments.
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pone-0069442-g007: The effect of AcMNPV infection on host gene activity.Genomic DNA from AcMNPV or mock -infected cells were digested with DNase I. The percentage of DNA resistant to the nuclease was determined with qPCR with primers for S. frugiperda β-Actin and β-Tubulin. The error bars indicate the standard deviation calculated from at least three independent parallel experiments.

Mentions: Whether Su(var) 3-9 is involved in AcMNPV infection and virus–host interaction is studied. AcMNPV-infected Sf9 cells (MOI=20 plaque-forming units per cell) were examined for Su(var) 3-9 transcription by qPCR. As shown in Figure 6, transcriptions of cellular cytoskeleton genes, β-Actin and β-Tubulin declined dramatically 8-12 h post infection (p.i.), whereas the mRNA level of Su(var) 3-9 increased to more than threefold higher than that of the control after 12 h p.i. The transcriptions level of HP1a and HP1b remain similar to those in the control. DNase I-sensitivity assay was executed to examine whether the abnormal expression trends of Su(var) 3-9, HP1a and HP1b, in contrast with the majority of host genes that decreased their expression after virus infection, caused chromatin compaction. The results showed that the cellular DNA of infected Sf9 cells became less sensitive to DNase I than that of mock infected (Figure 7) at the loci of β-Actin and β-Tubulin genes 24 h p.i.


Identification and characterization of the Spodoptera Su(var) 3-9 histone H3K9 trimethyltransferase and its effect in AcMNPV infection.

Li B, Li S, Yin J, Zhong J - PLoS ONE (2013)

The effect of AcMNPV infection on host gene activity.Genomic DNA from AcMNPV or mock -infected cells were digested with DNase I. The percentage of DNA resistant to the nuclease was determined with qPCR with primers for S. frugiperda β-Actin and β-Tubulin. The error bars indicate the standard deviation calculated from at least three independent parallel experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3722159&req=5

pone-0069442-g007: The effect of AcMNPV infection on host gene activity.Genomic DNA from AcMNPV or mock -infected cells were digested with DNase I. The percentage of DNA resistant to the nuclease was determined with qPCR with primers for S. frugiperda β-Actin and β-Tubulin. The error bars indicate the standard deviation calculated from at least three independent parallel experiments.
Mentions: Whether Su(var) 3-9 is involved in AcMNPV infection and virus–host interaction is studied. AcMNPV-infected Sf9 cells (MOI=20 plaque-forming units per cell) were examined for Su(var) 3-9 transcription by qPCR. As shown in Figure 6, transcriptions of cellular cytoskeleton genes, β-Actin and β-Tubulin declined dramatically 8-12 h post infection (p.i.), whereas the mRNA level of Su(var) 3-9 increased to more than threefold higher than that of the control after 12 h p.i. The transcriptions level of HP1a and HP1b remain similar to those in the control. DNase I-sensitivity assay was executed to examine whether the abnormal expression trends of Su(var) 3-9, HP1a and HP1b, in contrast with the majority of host genes that decreased their expression after virus infection, caused chromatin compaction. The results showed that the cellular DNA of infected Sf9 cells became less sensitive to DNase I than that of mock infected (Figure 7) at the loci of β-Actin and β-Tubulin genes 24 h p.i.

Bottom Line: Su(var) 3-9 protein was found to be localized in the nucleus in Sf9 cells, and interact with histone H3, and the heterochromatin protein 1a (HP1a) and HP1b.A dose-dependent enzymatic activity was found at both 27 °C and 37 °C in vitro, with higher activity at 27 °C.Addition of specific inhibitor chaetocin resulted in decreased histone methylation level and host chromatin relaxation.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, People's Republic of China.

ABSTRACT
Histone H3-lysine(9) (H3K9) trimethyltransferase gene Su(var) 3-9 was cloned and identified in three Spodoptera insects, Spodopterafrugiperda (S. frugiperda), S. exigua and S. litura. Sequence analysis showed that Spodoptera Su(var) 3-9 is highly conserved evolutionarily. Su(var) 3-9 protein was found to be localized in the nucleus in Sf9 cells, and interact with histone H3, and the heterochromatin protein 1a (HP1a) and HP1b. A dose-dependent enzymatic activity was found at both 27 °C and 37 °C in vitro, with higher activity at 27 °C. Addition of specific inhibitor chaetocin resulted in decreased histone methylation level and host chromatin relaxation. In contrast, overexpression of Su(var) 3-9 caused increased histone methylation level and cellular genome compaction. In AcMNV-infected Sf9 cells, the transcription of Su(var) 3-9 increased at late time of infection, although the mRNA levels of most cellular genes decreased. Pre-treatment of Sf9 cells with chaetocin speeded up viral DNA replication, and increased the transcription level of a variety of virus genes, whereas in Sf9 cells pre-transformed with Su(var) 3-9 expression vector, viral DNA replication slow down slightly. These findings suggest that Su(var) 3-9 might participate in the viral genes expression an genome replication repression during AcMNPV infection. It provided a new insight for the understanding virus-host interaction mechanism.

Show MeSH
Related in: MedlinePlus