Sputnik virophage disrupts the transcriptional regulation of its host giant virus

Sputnik virophages are small double-stranded DNA (dsDNA) viruses that replicate only inside host amoebae infected with giant dsDNA viruses, mimiviruses. Sputnik infection affects mimivirus replication, but their molecular interaction remains poorly understood. Here, we performed a time-course transcriptome analysis of Acanthamoeba castellanii cells infected with Acanthamoeba polyphaga mimivirus (APMV) (hereafter referred to as Sputnik ⁻ cells) and those infected with both APMV and Sputnik 3 virophage (Sputnik ⁺ cells). The gene expression patterns of the amoebae were similar between these two conditions, whereas the expression of APMV genes was drastically affected by Sputnik, depending on the timing of their expression. Early expressed APMV genes showed similar expression patterns in both conditions at the early stage of infection. However, at late stages, their expression levels remained higher in Sputnik ⁺ cells than in Sputnik ⁻ cells, suggesting a prolongation of early gene expression by Sputnik. Late expressed APMV genes showed lower expression at earlier stages in Sputnik ⁺ cells, but their expression levels reached or exceeded those in Sputnik ⁻ cells at later stages, indicating a delay of gene expression. Overall, our results demonstrated that Sputnik infection drastically alters the transcriptome of APMV rather than amoeba, likely by disturbing the transition from early to late stages of APMV infection.