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Dynamic redistribution of nuclear matrix proteins by adenovirus infection
1Department of Pathology, University of Ulsan College of Medicine, Seoul 138-736, Korea.
Abstract:
We analyzed the redistribution of nuclear proteins, PML protein, Ku70/Ku80, a putative spliceosome associated protein, pNMM102 and a nucleolar proliferating antigen, p120 after adenovirus 5 (Ad5) infection using immunofluorescent staining. These proteins remained after in situ fractionation. PML was located at irregular bars 6 h after infection from fine dots of uninfected cells. Distribution pattern of PML was not changed throughout the course of infection. Internal nuclear matrix network composed of Ku protein became much coarser in interphase cells at 12 h after infection. Ku protein was clumped at 18 h after infection, where EIA protein was colocalized. Speckles and interconnecting fibrils recognized by monoclonal antibody NMM102 disappeared early after infection, and reappeared at 18 h after infection in various patterns. The number and staining intensity of p120 containing domains increased markedly in early replication phase, and their shape became irregular with a few fine dots. A few Ad5 infected cells revealed diffuse nucleoplasmic as well as nucleolar p120 in late replication phase. Redistribution of four different nuclear matrix proteins by Ad5 infection indicates that the nuclear matrix is dynamically involved in gene expression.
Insights
Adenovirus 5 infection causes significant nuclear matrix protein redistribution. Key proteins like PML, Ku70/Ku80, pNMM102, and p120 change localization, indicating dynamic nuclear matrix involvement in gene expression.
Area of Science:
- Cell Biology
- Virology
- Molecular Biology
Background:
- The nuclear matrix is a dynamic structure within the cell nucleus.
- Viral infections can alter host cell nuclear organization.
- Understanding nuclear matrix protein dynamics during viral infection is crucial for comprehending viral pathogenesis and host-pathogen interactions.
Purpose of the Study:
- To investigate the dynamic redistribution of specific nuclear matrix proteins following Adenovirus 5 (Ad5) infection.
- To elucidate the role of the nuclear matrix in Ad5-mediated gene expression.
Main Methods:
- Immunofluorescent staining was employed to visualize and analyze the localization of nuclear proteins.
- In situ fractionation techniques were used to confirm the presence of proteins within the nuclear matrix.
- Confocal microscopy was utilized to observe changes in protein distribution patterns over time post-infection.
Main Results:
- Adenovirus 5 infection induced significant redistribution of PML protein, Ku70/Ku80, pNMM102, and p120.
- PML protein shifted from fine dots to irregular bars post-infection.
- Ku protein network coarsened and later clumped, colocalizing with EIA protein.
- pNMM102 speckles disappeared early and reappeared in varied patterns.
- p120 containing domains increased in number and altered shape, with some cells showing diffuse nucleoplasmic and nucleolar localization.
Conclusions:
- The nuclear matrix undergoes dynamic changes in response to Ad5 infection.
- Redistribution of these nuclear matrix proteins suggests their active involvement in viral gene expression processes.
- These findings highlight the nuclear matrix as a key player in the interplay between viruses and host cell machinery.