Human papillomavirus infection requires the TSG101 component of the ESCRT machinery

•We show that the ESCRT component TSG101 is required for optimal infection with both HPV-16 and BPV-1.•We find that L2 capsid proteins from many Papillomavirus types can interact with TSG101.•We show that interaction between L2 and TSG101 alters the subcellular distribution of L2.•TSG101 can modulate the levels of L2 polyubiquitination.

Infection with human papillomaviruses (HPV) requires the minor capsid component L2, which plays an essential role in directing appropriate endosomal trafficking. Previous studies have indicated an infection route involving multi-vesicular bodies (MVBs), and an essential element in their biogenesis is the ESCRT machinery. Here we show that the ESCRT component TSG101 is required for optimal infection with both HPV-16 and BPV-1, with loss of TSG101 resulting in a decrease in viral infection, whereas overexpressed TSG101 increases rates of infection. We find that L2 proteins from multiple PV types interact with TSG101 and show that this interaction contributes to an alteration in the subcellular distribution of L2. In addition, TSG101 can modulate the levels of L2 polyubiquitination. These results demonstrate that TSG101 plays an important part in infection with diverse PVs, and suggests that trafficking of HPV through the ESCRT machinery and MVBs is part of infectious virus entry.