Summary
In rabbit kidney epithelial cells (RK13) abortively infected with human cytomegalovirus (HCMV), DNA synthesis at 1 or 2 days post-infection was enhanced 4 to 5 fold, compared to mock-infected cells. DNA analysis by isopycnic centrifugation revealed that the DNA newly synthesized in the virus infected RK13 cells was of cellular origin. HCMV infection also caused a marked increase in the mitotic activity of RK13 cells. When semi-confluent RK13 cells were infected more than 20 per cent of cells demonstrated mitosis at 72 hours post-infection although the rate of cell growth was considerably reduced compared to that of uninfected cells. The most frequent chromosomal change observed was fragmentation although other aberrations, gap, break, deletion etc. occurred also.
Two immediate-early viral polypeptides with apparent molecular weights 72,000 (72K) and 76,000 (76K) daltons were produced in both RK13 cells and human embryonic lung cells (HEL) by 3 hours post-infection. Synthesis of the 76K polypeptide was greater than that of the 72K polypeptide in non-permissive RK13 cells whereas the reverse occurred in permissive HEL cells. Furthermore, of three early polypeptides which were expressed in productively infected HEL cells two, 88K and 80K, were not detected in abortively infected RK13 cells.
These results suggest that the arrest in mitosis of the abortively infected RK13 cells and the subsequent chromosomal changes are associated with the altered expression of immediate-early or early virus functions in these cells.
Following HCMV infection permissive HEL cells undergo a characteristic sequence of morphological changes (3) of which cell rounding and “contraction” are observed in the early stages and could be related to either the action of cellular contractile proteins or changes in the plasma membrane permeability and a concomitant Ca++ influx (2). ‘Relaxation’ and cell enlargement are other changes considered to be initiated by early virus functions although they occur after the commencement of virus DNA synthesis. Thus, the expression of immediate-early and early virus functions has an important role in controlling the sequential morphological changes in infected cells and may also be important in determining whether infection with HCMV will be productive or abortive.
Similar content being viewed by others
References
Albrecht, T., Nachtigal, M., St. Jeor, S. C., Rapp, F.: Induction of cellular DNA synthesis and increased mitotic activity in syrian hamster embryo cells abortively infected with human cytomegalovirus. J. gen. Virol.30, 167–177 (1976).
Albrecht, T., Speelman, D. J., Steinsland, O. S.: Similarities between cytomegalovirus-induced cell rounding and contraction of smooth muscle cells. Life Sci.32, 2273–2278 (1983).
Albrecht, T., Li, J. L., Speelman, D., Ball, B., Nokta, M., Fons, M., Lee, C. H., Steinsland, O., Thompson, C. W., Carhey, D. H.: Cellular responses to human cytomegalovirus infection. In:Plotkin, S. A., Michelson, S., Pagano, J., Rapp, F. (eds.), CMV: Pathogenesis and Prevention of Human Infection, Birth Defects. Original Article Series, Vol. 20, No. 1, 21–34. New York: Alan R. Liss 1984.
Blanton, R. A., Tevethia, M. J.: Immunoprecipitation of virus-specific immediate-early and early polypeptides from cells lytically infected with human cytomegalovirus strain AD 169. Virology112, 262–273 (1981).
Boldogh, I., Gönczöl, É., Gärtner, L., Váczi, L.: Stimulation of host DNA synthesis and induction of early antigens by ultraviolet light irradiated human cytomegalovirus. Arch. Virol.58, 289–299 (1978).
Crouch, N. A., Rapp, F.: Cell-dependent differences in the production of infectious herpes simplex virus at a supraoptimal temperature. J. Virol.9, 223–230 (1970).
DeMarchi, J. M., Kaplan, A. S.: The role of defective cytomegalovirus particles in the induction of host cell DNA synthesis. Virology82, 93–99 (1977).
DeMarchi, J. M.: Correlation between stimulation of host cell DNA synthesis by human cytomegalovirus and lack of expression of a subset of early virus genes. Virology129, 274–286 (1983).
DeMarchi, J. M.: Nature of the block in the expression of some early virus genes in cells abortively infected with human cytomegalovirus. Virology129, 287–297 (1983).
Epstein, A. L., Jacquemont, B.: Virus polypeptide synthesis induced by herpes simplex virus in non-permissive rat XC cells. J. gen. Virol.64, 1499–1508 (1983).
Färber, I., Wutzler, P., Schweizer, H., Sprossig, M.: Human cytomegalovirus induced changes in rabbit cells. Arch. Virol.59, 257–261 (1979).
Fioretti, A., Furukawa, T., Santoli, D., Plotkin, S. A.: Nonproductive infection of guinea pig cells with human cytomegalovirus. J. Virol.11, 998–1003 (1973).
Fried, M., Pitts, J. D.: Replication of polyoma virus DNA. I. A resting cell system for biochemical studies on polyoma virus. Virology34, 761–770 (1968).
Furukawa, T., Tanaka, S., Plotkin, S. A.: Stimulation of macromolecular synthesis in guinea pig cells by human CMV. Proc. Soc. Exp. Biol. Med.148, 211–214 (1975).
Gönczöl, É., Plotkin, S. A.: Cells infected with human cytomegalovirus release a factor(s) that stimulates cell DNA synthesis. J. gen. Virol.65, 1833–1837 (1984).
Hamper, B., Ellison, S. A.: Chromosomal aberrations induced by an animal virus. Nature (London)192, 145–146 (1961).
Hart, H., Norval, M.: Association of human cytomegalovirus (HCMV) with mink and rabbit lung cells. Arch. Virol.67, 203–215 (1981).
Hirai, K., Furukawa, T., Plotkin, S. A.: Induction of DNA polymerase in WI-38 and guinea pig cells infected with human cytomegalovirus (HCMV). Virology70, 251–255 (1976).
Jacquemont, B., Verrier, B., Epstein, A. L., Machuca, I.: Expression of immediate-early genes in herpes simplex virus type 1-infected XC cells: Lack of ICP 22 (68K) polypeptide. J. gen. Virol.65, 1331–1340 (1984).
Kamiya, S., Tanaka, J., Ogura, T., Sato, H., Ogura, H., Yoshie, T., Hatano, M.: Abortive infection with human cytomegalovirus induces an alteration of growth pattern: Morphological changes with cytocidal effect in rabbit kidney epithelial cells. Arch. Virol.86, 143–150 (1985).
Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London)227, 680–685 (1970).
Lüleoí, G., Sakízlí, M., Günalp, A.: Selective chromosomal damage caused by human cytomegalovirus. Acta. Virol.24, 341–345 (1980).
Michelson, S., Horodniceanu, F., Kress, M., Tardy-Panit, M.: Human cytomegalovirus-induced immediate early antigens: Analysis in sodium dodecyl sulfate-polyacrylamide gel electrophoresis after immunoprecipitation. J. Virol.32, 259–267 (1979).
Moorehead, P. S., Nowell, P. C., Mellman, W. J., Battips, D. M., Hingerford, D. A.: Chromosome preparations of leukocytes cultured from human peripheral blood. Exp. Cell Res.20, 613–616 (1960).
Nachtigal, M., Nachtigal, S.: Interactions between human herpesviruses and host cell chromosomes. Arch. Roum. Path. Exp. Microbiol.37, 223–249 (1978).
Ogura, H., Sato, H., Tanaka, J., Kamiya, S., Yoshie, T., Hatano, M.: Synthesis of M protein of HVJ (Sendai virus) in rat glial cells is selectively restricted at a non-permissive temperature. J. gen. Virol.65, 639–643 (1984).
O'Neill, F. J., Miles, C. P.: Chromosome changes in human cells induced by herpes simplex, type 1 and 2. Nature (London)223, 851–852 (1969).
O'Neill, F. J., Rapp, F.: Early events required for induction of chromosome abnormalities in human cells by herpes simplex virus. Virology44, 544–553 (1971).
Stinski, M. F.: Human cytomegalovirus: glycoproteins associated with virions and dense bodies. J. Virol.19, 594–609 (1976).
Stinski, M.: Sequence of protein synthesis in cells infected by human cytomegalovirus: Early and late virus-induced polypeptides. J. Virol.26, 686–701 (1978).
St. Jeor, S. C., Albrecht, T., Funk, F. D., Rapp, F.: Stimulation of cellular DNA synthesis by human cytomegalovirus. J. Virol.13, 353–362 (1974).
Tanaka, J., Ogura, T., Kamiya, S., Sato, H., Yoshie, T., Ogura, H., Hatano, M.: Enhanced replication of human cytomegalovirus in human fibroblast treated with dexamethasone. J. gen. Virol.65, 1759–1767 (1984).
Waner, J. L., Weller, T. H.: Behavior of human cytomegaloviruses in cell cultures of bovine and simian origin. Proc. Soc. Exp. Biol. Med.145, 379–384 (1974).
Wentworth, B. B., French, L.: Plaque assay of cytomegalovirus strains of human origin. Proc. Soc. Exp. Biol. Med.135, 253–258 (1970).
Yamanishi, K., Rapp, F.: Induction of host DNA synthesis and DNA polymerase by DNA-negative temperature-sensitive mutants of human cytomegalovirus. Virology94, 237–241 (1979).
Author information
Authors and Affiliations
Additional information
With 4 Figures
Rights and permissions
About this article
Cite this article
Kamiya, S., Tanaka, J., Ogura, T. et al. Rabbit kidney cells abortively infected with human cytomegalovirus are arrested in mitotic phase. Archives of Virology 89, 131–144 (1986). https://doi.org/10.1007/BF01309884
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01309884