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Isolation of Viral Replication Compartment-enriched Sub-nuclear Fractions from Adenovirus-infected Normal Human Cells
Published on: November 12, 2015
Alterations in nuclear matrix structure after adenovirus infection
Abstract:
Infection of HeLa cells with adenovirus serotype 2 causes rearrangements in nuclear matrix morphology which can best be seen by gentle cell extraction and embedment-free section electron microscopy. We used these techniques to examine the nuclear matrices and cytoskeletons of cells at 6, 13, 28, and 44 h after infection. As infection progressed, chromatin condensed onto the nucleoli and the nuclear lamina. Virus-related inclusions appeared in the nucleus, where they partitioned with the nuclear matrix. These virus centers consisted of at least three distinguishable areas: amorphously dense regions, granular regions whose granulations appeared to be viral capsids, and filaments connecting these regions to each other and to the nuclear lamina. The filaments became decorated with viral capsids of two different densities, which may be empty capsid shells and capsids with DNA-protein cores. The interaction of some capsids with the filaments persisted even after lysis of the cell. We propose that granulated virus-related structures are sites of capsid assembly and storage and that the filaments may be involved in the transport of capsids and capsid intermediates. The nuclear lamina became increasingly crenated after infection, with some extensions appearing to bud off and form blebs of nuclear material in the cytoplasm. The perinuclear cytoskeleton became rearranged after infection, forming a corona of decreased filament number around the nucleus. In summary, we propose that adenovirus rearranges the nuclear matrix and cytoskeleton to support its own replication.
Insights
Adenovirus infection rearranges HeLa cell nuclear structures, forming virus assembly sites and altering the cytoskeleton. These changes support viral replication by reorganizing the nuclear matrix and cytoskeleton.
Area of Science:
- Cell Biology
- Virology
- Microscopy
Background:
- Adenovirus serotype 2 infection impacts host cell nuclear organization.
- Understanding these nuclear rearrangements is crucial for viral replication mechanisms.
Purpose of the Study:
- To investigate the morphological changes in nuclear matrix and cytoskeleton of HeLa cells post-adenovirus infection.
- To elucidate the role of nuclear structures in adenovirus replication.
Main Methods:
- Gentle cell extraction and embedment-free section electron microscopy were employed.
- HeLa cells were analyzed at various time points (6, 13, 28, 44 h) after infection.
Main Results:
- Chromatin condensed, and virus-related inclusions formed within the nuclear matrix.
- Filaments within virus centers were decorated with viral capsids, suggesting assembly and transport roles.
- Nuclear lamina became crenated, and the perinuclear cytoskeleton rearranged.
Conclusions:
- Adenovirus infection induces significant rearrangements of the nuclear matrix and cytoskeleton.
- Proposed that nuclear structures facilitate viral capsid assembly, storage, and transport.
- Nuclear and cytoskeletal alterations are essential for supporting adenovirus replication.
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