Alterations in mouse mammary tumor virus (MMTV) production and composition were induced by exposure of mammary tumor cells to cytodisruptive agents. Treatment with 2.1 microM cytochalasin D (CD) for 24 h reduced MMTV yield by 80% and electron microscopic examination of these cells did not reveal budding virions. Immune precipitation and quantitative immunofluorescence studies demonstrated that CD had no significant effect on MMTV polypeptide synthesis or surface expression suggesting that CD inhibited late steps in MMTV maturation. Decreases in MMTV production were also observed as a result of 24 h exposure of the cells to 2.1 microM cytochalasin B (CB). However, an initial 70% increase in the levels of extracellular virions within the first 18 h of treatment preceded diminution of virus production. In addition, CB was unable to abrogate maturation and release of MMTV particles as revealed by electron microscopic evaluation of thin sections of treated cells. Colcemid at 0.28 microM had no effect on virus production during the first 24 h of exposure although MMTV yield was reduced by 60-70% after 36 h of treatment. Polypeptide profiles of MMTV purified from cell cultures treated with any of the three cytodisruptive agents were altered and included 5-7 polypeptides not typically present in MMTV from untreated cells. These cytodisruptive agents did not significantly affect viability and protein metabolism of MJY-alpha cells; the data suggest that alterations in MMTV replication were due to disruption of the cellular cytoskeleton.