Bovine papillomavirus E1 protein affects the host cell cycle phase fractions

Abstract

C127 murine fibroblast cells were electroporated with a bovine papillomavirus E1 protein expression vector and examined by flow cytometry. E1 expressing cells (E1+) within the total cell population were distinguished from nonexpressing cells (E1-) by immunofluorescent staining with anti-E1 serum and a fluorescein-conjugated second antibody. Under conditions of saturation with the first and second antibodies, the specific green fluorescence reflected the level of intracellular E1 protein. Simultaneous staining with a DNA-specific dye, propidium iodide (PI), enabled the cell cycle distributions for the E1+ and E1- cell populations to be determined. It was found that the E1+ subpopulation had a reduced percentage of cells in G1 phase and an increased percentage of G2+M phase cells, compared to the E1- subpopulation. There was no significant difference in overall doubling time or percentage of noncycling cells in the E1+ vs. E1- populations, indicating that the change in cell cycle distribution was not due to a general activation or inhibition of cell growth by E1. Direct measurement of cell cycle phase fractions confirmed that the G1 phase was decreased and the G2+M phase was increased in E1 expressing cells. As these observations were made in the absence of other viral proteins or viral DNA replication, it suggests that the E1 protein exerts an effect on the host cell independent of its direct role in viral DNA replication. Thus, E1 may interact directly with the host cell cycle regulatory machinery.