Involvement of microtubules in cytopathic effects of animal viruses: early proteins of adenovirus and herpesvirus inhibit formation of microtubular paracrystals in HeLa-S3 cells.

In order to examine the involvement of microtubules in the virus-induced cytopathic effect (c.p.e.), the effect of virus infection on the formation of microtubular paracrystals (PC) induced by 10 microgram/ml of vinblastine sulphate in HeLa-S3 cells was examined by phase-contrast microscopy. In poliovirus-infected cells, c.p.e. (cell rounding) and the inhibition of PC formation proceeded in parallel, starting 4 h post-infection. In Sendai virus-infected cells, however, PC formation was not inhibited even 24 h post-infection when most infected cells clearly showed c.p.e. (syncytial formation). In adenovirus-infected cells, the inhibition of PC formation was observed 9 h before the appearance of c.p.e. Cytosine arabinoside (ara C) did not block the inhibition of PC formation in infected cells, but blocked the appearance of late c.p.e. (nuclear alteration). Cycloheximide blocked both the inhibition of PC formation and the induction of late c.p.e. These results suggest that an early protein synthesized de novo by adenovirus is required for direct or indirect inhibition of the microtubular PC formation. Furthermore, on ultraviolet (u.v.) inactivation of adenovirus both activities (induction of early c.p.e. shown by shrinkage of cytoplasm, and inhibition of PC formation) followed the same inactivation curve and were inactivated at a slower rate than viral infectivity and the activity leading to late c.p.e. The u.v. light sensitive target responsible for the induction of early c.p.e. and the inhibition of PC formation is about 20% of that for infectivity and is in accord with the genome size of the early functioning virus genes. In herpes simplex virus (HSV)-infected cells, the inhibition of PC formation, the appearance of c.p.e. (cell rounding and disappearance of nucleoli) and the synthesis of V antigen proceeded in parallel. These three functions of HSV were not blocked in infected cells even when the de novo synthesis of virus DNA was inhibited by ara C or phosphonoacetic acid (PAA), whereas these three functions were blocked by cycloheximide, suggesting that a protein coded by the input virus genome early after infection inhibits the microtubular PC formation and is responsible for c.p.e. From the u.v. inactivation curve of HSV, it was confirmed that only one-tenth of virus genome was responsible for both activities (induction of c.p.e. and inhibition of PC formation).