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HDAC inhibitors stimulate viral transcription by multiple mechanisms.

Balakrishnan L, Milavetz B - Virol. J. (2008)

Bottom Line: The effects of histone deacetylase inhibitor (HDACi) treatment on SV40 transcription and replication were determined by monitoring the levels of early and late expression, the extent of replication, and the percentage of SV40 minichromosomes capable of transcription and replication following treatment with sodium butyrate (NaBu) and trichostatin A (TSA).The HDACi treatment was found to maximally stimulate early transcription at early times and late transcription at late times through increased numbers of minichromosomes which carry RNA polymerase II (RNAPII) transcription complexes and increased occupancy of the transcribing minichromosomes by RNAPII.The increased recruitment of transcribing minichromosomes at late times was correlated to a corresponding reduction in SV40 replication and the percentage of minichromosomes capable of replication.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, University of North Dakota, Grand Forks, North Dakota, USA. lbalakrishnan@medicine.nodak.edu

ABSTRACT

Background: The effects of histone deacetylase inhibitor (HDACi) treatment on SV40 transcription and replication were determined by monitoring the levels of early and late expression, the extent of replication, and the percentage of SV40 minichromosomes capable of transcription and replication following treatment with sodium butyrate (NaBu) and trichostatin A (TSA).

Results: The HDACi treatment was found to maximally stimulate early transcription at early times and late transcription at late times through increased numbers of minichromosomes which carry RNA polymerase II (RNAPII) transcription complexes and increased occupancy of the transcribing minichromosomes by RNAPII. HDACi treatment also partially relieved the normal down-regulation of early transcription by T-antigen seen later in infection. The increased recruitment of transcribing minichromosomes at late times was correlated to a corresponding reduction in SV40 replication and the percentage of minichromosomes capable of replication.

Conclusion: These results suggest that histone deacetylation plays a critical role in the regulation of many aspects of an SV40 lytic infection.

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Effects of HDACi treatment on chromosomes carrying RNA Polymerase II. 30 min, 8 hour and 48 hour SV40 minichromosomes either untreated or treated with 250 μM NaBu or 120 nM TSA were immune selected using 10 μl of antibody against RNAPII. Data is expressed as the percentage of SV40 minichromosomes containing RNA Polymerase II and represent the average of three independent experiments.
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Figure 2: Effects of HDACi treatment on chromosomes carrying RNA Polymerase II. 30 min, 8 hour and 48 hour SV40 minichromosomes either untreated or treated with 250 μM NaBu or 120 nM TSA were immune selected using 10 μl of antibody against RNAPII. Data is expressed as the percentage of SV40 minichromosomes containing RNA Polymerase II and represent the average of three independent experiments.

Mentions: SV40 minichromosomes from control and HDACi treated infected cells were harvested in parallel at 30 minutes, 8 hours, and 48 hours post-infection from at least three separate preparations of SV40 minichromosomes at each time point and subjected to a ChIP analysis with antibody to RNAPII followed by real-time PCR amplification for quantitation. From the CT values obtained from the analyses of HDACi treated and untreated SV40 minichromosomes we then determined the percentage of the SV40 minichromosomes that contained bound RNAPII from each set of samples (Figure 2).


HDAC inhibitors stimulate viral transcription by multiple mechanisms.

Balakrishnan L, Milavetz B - Virol. J. (2008)

Effects of HDACi treatment on chromosomes carrying RNA Polymerase II. 30 min, 8 hour and 48 hour SV40 minichromosomes either untreated or treated with 250 μM NaBu or 120 nM TSA were immune selected using 10 μl of antibody against RNAPII. Data is expressed as the percentage of SV40 minichromosomes containing RNA Polymerase II and represent the average of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2291040&req=5

Figure 2: Effects of HDACi treatment on chromosomes carrying RNA Polymerase II. 30 min, 8 hour and 48 hour SV40 minichromosomes either untreated or treated with 250 μM NaBu or 120 nM TSA were immune selected using 10 μl of antibody against RNAPII. Data is expressed as the percentage of SV40 minichromosomes containing RNA Polymerase II and represent the average of three independent experiments.
Mentions: SV40 minichromosomes from control and HDACi treated infected cells were harvested in parallel at 30 minutes, 8 hours, and 48 hours post-infection from at least three separate preparations of SV40 minichromosomes at each time point and subjected to a ChIP analysis with antibody to RNAPII followed by real-time PCR amplification for quantitation. From the CT values obtained from the analyses of HDACi treated and untreated SV40 minichromosomes we then determined the percentage of the SV40 minichromosomes that contained bound RNAPII from each set of samples (Figure 2).

Bottom Line: The effects of histone deacetylase inhibitor (HDACi) treatment on SV40 transcription and replication were determined by monitoring the levels of early and late expression, the extent of replication, and the percentage of SV40 minichromosomes capable of transcription and replication following treatment with sodium butyrate (NaBu) and trichostatin A (TSA).The HDACi treatment was found to maximally stimulate early transcription at early times and late transcription at late times through increased numbers of minichromosomes which carry RNA polymerase II (RNAPII) transcription complexes and increased occupancy of the transcribing minichromosomes by RNAPII.The increased recruitment of transcribing minichromosomes at late times was correlated to a corresponding reduction in SV40 replication and the percentage of minichromosomes capable of replication.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, University of North Dakota, Grand Forks, North Dakota, USA. lbalakrishnan@medicine.nodak.edu

ABSTRACT

Background: The effects of histone deacetylase inhibitor (HDACi) treatment on SV40 transcription and replication were determined by monitoring the levels of early and late expression, the extent of replication, and the percentage of SV40 minichromosomes capable of transcription and replication following treatment with sodium butyrate (NaBu) and trichostatin A (TSA).

Results: The HDACi treatment was found to maximally stimulate early transcription at early times and late transcription at late times through increased numbers of minichromosomes which carry RNA polymerase II (RNAPII) transcription complexes and increased occupancy of the transcribing minichromosomes by RNAPII. HDACi treatment also partially relieved the normal down-regulation of early transcription by T-antigen seen later in infection. The increased recruitment of transcribing minichromosomes at late times was correlated to a corresponding reduction in SV40 replication and the percentage of minichromosomes capable of replication.

Conclusion: These results suggest that histone deacetylation plays a critical role in the regulation of many aspects of an SV40 lytic infection.

Show MeSH
Related in: MedlinePlus