ADAM17-dependent signaling is required for oncogenic human papillomavirus entry platform assembly.

Karina Reiss,Fatima Boukhallouk,Snježana Mikuličić,Dominik Sons,Jérôme Finke,Luise Florin,Thorsten Lang,Yahya Homsi,Elena Wüstenhagen

doi:10.7554/elife.44345

Abstract

Oncogenic human papillomaviruses (HPV) are small DNA viruses that infect keratinocytes. After HPV binding to cell surface receptors, a cascade of molecular interactions mediates the infectious cellular internalization of virus particles. Aside from the virus itself, important molecular players involved in virus entry include the tetraspanin CD151 and the epidermal growth factor receptor (EGFR). To date, it is unknown how these components are coordinated in space and time. Here, we studied plasma membrane dynamics of CD151 and EGFR and the HPV16 capsid during the early phase of infection. We find that the proteinase ADAM17 activates the extracellular signal-regulated kinases (ERK1/2) pathway by the shedding of growth factors which triggers the formation of an endocytic entry platform. Infectious endocytic entry platforms carrying virus particles consist of two-fold larger CD151 domains containing the EGFR. Our finding clearly dissects initial virus binding from ADAM17-dependent assembly of a HPV/CD151/EGFR entry platform.

Highlights

Viral infections by human papillomaviruses (HPVs) cause benign warts and malignant tumors
Infecting HaCaT keratinocytes with HPV16 pseudovirions (PsVs), we applied the broad-spectrum metalloproteinase inhibitor TAPI-0, the preferential ADAM10 inhibitor GI254023X (GI) and the mixed ADAM10/ADAM17 inhibitor GW280264X (GW) (Figure 1—figure supplement 1A)
For ADAM10 and ADAM17, respectively, two different siRNAs were employed in three epithelial cell models, including human cervical carcinoma cell line (HeLa) cells, HaCaT cells, and primary normal human epithelial keratinocytes (NHEK)

Summary

Introduction

Viral infections by human papillomaviruses (HPVs) cause benign warts and malignant tumors. More than 240 papillomavirus types have been characterized in diverse hosts, including mammals, birds and reptiles. The oncogenic HPV types 16, 18, and 31 are responsible for severe human cancers, including cervical cancer and anogenital, head and neck tumors (Doorbar et al, 2012). HPV infections require a micro-wound enabling virion binding to mitotically active basal keratinocytes (Doorbar et al, 2012; Ozbun, 2019). The long halftime points toward a complex cascade of events at the cell membrane prior to virus entry into host cells. It is assumed that the sequence is initiated upon virus binding to a primary receptor, from which the virus is released in a modified form, followed by rebinding to a secondary receptor complex, which is endocytosed (Ozbun, 2019; Raff et al, 2013; Mikulicicand Florin, 2019)

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