Nested parasitic chains are common schemes in nature, not limited to cellular organisms. Some giant viruses infecting protists are hyperparasitized by smaller viruses named virophages. Both can carry episomal plasmid-like DNA molecules known as transpovirons in their particles. They all share common transcriptional regulatory elements dictating the expression of their genes, which are transcribed within viral factories built by giant viruses in the host cytoplasm. This suggests close but as yet undetermined interactions between their respective transcriptional networks. Here, we studied the protein content of Megavirus chilensis virions produced in Acanthamoeba castellanii cells co-infected or not with the virophage Zamilon vitis.