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Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins

Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins

Published on: August 31, 2017

Herpes simplex virus induces a processing factor that stimulates poly(A) site usage.

J McLauchlan¹, S Simpson, J B Clements

¹MRC Virology Unit, University of Glasgow, Scotland.

|December 22, 1989

Summary

This summary is machine-generated.

Nuclear extracts from herpes simplex virus-infected cells show increased RNA processing at a specific viral poly(A) site. This heat-labile activity, observed in vitro and in vivo, enhances late viral polyadenylation efficiency.

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Temporal Analysis of the Nuclear-to-cytoplasmic Translocation of a Herpes Simplex Virus 1 Protein by Immunofluorescent Confocal Microscopy

Temporal Analysis of the Nuclear-to-cytoplasmic Translocation of a Herpes Simplex Virus 1 Protein by Immunofluorescent Confocal Microscopy

Published on: November 4, 2018

Area of Science:

Molecular Biology
Virology
Gene Expression

Background:

Herpes simplex virus (HSV) gene expression involves post-transcriptional modifications, including polyadenylation.
Polyadenylation site selection is a critical regulatory step in RNA processing.
Understanding viral RNA processing factors can reveal mechanisms of viral gene regulation.

Purpose of the Study:

To investigate the processing of RNA at herpes simplex virus poly(A) sites.
To identify potential cellular or viral factors influencing HSV polyadenylation efficiency.
To determine if specific viral poly(A) sites are processed differently in infected cells.

Main Methods:

In vitro processing assays using nuclear extracts from HSV-infected and mock-infected cells.
Comparison of poly(A) site processing efficiency using precursor RNAs with tandem viral poly(A) sites.
Heat lability assays to characterize the identified processing activity.
Analysis of viral RNAs from recombinant viruses to confirm in vivo findings.

Main Results:

Nuclear extracts from HSV-infected cells exhibit enhanced processing activity at a specific late HSV poly(A) site.
A second HSV poly(A) site showed similar processing efficiency in both infected and mock-infected cell extracts.
The observed specific processing activity was found to be heat labile.
In vivo studies using recombinant viruses corroborated the increased processing at the late viral poly(A) site.

Conclusions:

A specific, heat-labile activity present in HSV-infected cells enhances the processing of a late viral poly(A) site.
This factor plays a role in regulating polyadenylation efficiency at specific HSV poly(A) sites.
The findings suggest a mechanism for differential regulation of viral gene expression during HSV infection.