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Isolation of Viral Replication Compartment-enriched Sub-nuclear Fractions from Adenovirus-infected Normal Human Cells
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Precursor of human adenovirus core polypeptide Mu targets the nucleolus and modulates the expression of E2 proteins

T W R Lee1, F J Lawrence2, V Dauksaite3

  • 1School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.

The Journal of General Virology
|January 14, 2004
PubMed

Insights

Human adenovirus 2 (HAdV-2) preMu protein localizes to the nucleolus and inhibits late gene expression by affecting E2 protein accumulation. This suggests preMu modulates adenovirus E2 region protein accumulation.

Area of Science:

  • Molecular Biology
  • Virology
  • Cell Biology

Background:

  • Human adenovirus 2 (HAdV-2) is a significant human pathogen.
  • Understanding adenovirus protein function and localization is crucial for antiviral strategies.

Purpose of the Study:

  • To investigate the subcellular localization of HAdV-2 preMu and mature Mu (pX) proteins.
  • To determine the role of preMu in adenovirus replication and gene expression.

Main Methods:

  • Fusion proteins of preMu and Mu with enhanced green fluorescence protein (EGFP) were constructed.
  • Subcellular localization was analyzed using fluorescence microscopy in infected and transiently transfected cells.
  • Deletion mutagenesis was employed to identify functional domains of preMu.

Main Results:

  • preMu-EGFP exclusively localized to the nucleolus, containing a single nucleolar accumulation signal.
  • Both preMu-EGFP and Mu-EGFP were excluded from adenovirus DNA-binding protein (DBP)-rich replication centers.
  • Transient expression of preMu-EGFP, but not Mu-EGFP, prior to or during Ad2 infection inhibited late adenovirus gene expression.
  • This inhibition was linked to a failure in pre-terminal protein (preTP) expression from the E2 region, despite DBP expression.
  • A conserved C-terminal region of preMu was identified as responsible for these effects.

Conclusions:

  • preMu is a nucleolar protein that can inhibit adenovirus late gene expression.
  • preMu appears to modulate the accumulation of proteins from the adenovirus E2 region, potentially impacting preTP levels.
  • These findings suggest a novel regulatory role for preMu in the adenovirus life cycle.

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