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Organization of cytoskeleton elements during herpes simplex virus type 1 infection of human fibroblasts: an
Abstract:
Cultured human fibroblasts showed a typical fibrillar organization of microtubules in immunofluorescence, including the vimentin type of intermediate filament as well as actin-containing microfilaments. During infection with herpes simplex virus type 1 (HSV-1), the vimentin organization was maintained whereas actin, myosin and tubulin showed a progressive association with the viral glycoproteins within juxtanuclear structures. These structures could also be revealed with fluorochrome-coupled wheat germ agglutinin. Disruption of the microtubules by demecolcine treatment or their stabilization by taxol treatment did not prevent the aggregation of viral proteins in the cytoplasm. Taxol stabilization of the microtubules allowed the juxtanuclear accumulation of the glycoproteins in HSV-infected cells whereas treatment with demecolcine led to an accumulation of the glycoproteins either in small vesicles in the cytoplasm or in the focal adhesion areas of the cells. Production of infectious intracellular virus particles was reduced in cells treated with demecolcine or with taxol before and during infection. The results of this study indicate that the normal intracellular transport and distribution of the HSV glycoproteins and the formation of infectious virus are dependent on the presence of intact microtubules.
Insights
Herpes simplex virus type 1 (HSV-1) infection disrupts cellular transport. Intact microtubules are essential for proper glycoprotein distribution and infectious virus production in fibroblasts.
Area of Science:
- Cell Biology
- Virology
- Microscopy
Background:
- Cultured human fibroblasts exhibit organized microtubules, vimentin intermediate filaments, and actin microfilaments.
- Herpes simplex virus type 1 (HSV-1) infection is a common human viral infection.
Purpose of the Study:
- To investigate the role of cellular cytoskeleton, specifically microtubules, in the intracellular transport and assembly of HSV-1 glycoproteins.
- To determine the impact of microtubule disruption or stabilization on HSV-1 glycoprotein localization and infectious virus production.
Main Methods:
- Immunofluorescence microscopy was used to visualize microtubules, vimentin, actin, myosin, and tubulin.
- Treatment with demecolcine (microtubule disruptor) and taxol (microtubule stabilizer) was applied to HSV-1 infected fibroblasts.
- Wheat germ agglutinin was used to identify juxtanuclear structures.
Main Results:
- HSV-1 infection caused actin, myosin, and tubulin to associate with viral glycoproteins in juxtanuclear structures.
- Microtubule disruption or stabilization did not prevent viral protein aggregation but altered glycoprotein localization.
- Demecolcine treatment led to cytoplasmic vesicles or focal adhesion accumulation, while taxol promoted juxtanuclear accumulation of glycoproteins.
- Infectious virus particle production was reduced in cells treated with demecolcine or taxol.
Conclusions:
- Intact microtubules are crucial for the normal intracellular transport and distribution of HSV glycoproteins.
- The formation of infectious HSV particles depends on the integrity of the microtubule network within host cells.