Faculty Collaboration Database

Low-affinity binding in cis to P2Y₂R mediates force-dependent integrin activation during hantavirus infection. Mol Biol Cell 2017 Oct 15;28(21):2887-2903

Date

08/25/2017

Pubmed ID

28835374

Pubmed Central ID

PMC5638590

DOI

10.1091/mbc.E17-01-0082

Scopus ID

2-s2.0-85031296470 (requires institutional sign-in at Scopus site)   12 Citations

Abstract

Pathogenic hantaviruses bind to the plexin-semaphorin-integrin (PSI) domain of inactive, β₃ integrins. Previous studies have implicated a cognate cis interaction between the bent conformation β₅/β₃ integrins and an arginine-glycine-aspartic acid (RGD) sequence in the first extracellular loop of P2Y₂R. With single-molecule atomic force microscopy, we show a specific interaction between an atomic force microscopy tip decorated with recombinant α_IIbβ₃ integrins and (RGD)P2Y₂R expressed on cell membranes. Mutation of the RGD sequence to RGE in the P2Y₂R removes this interaction. Binding of inactivated and fluorescently labeled Sin Nombre virus (SNV) to the integrin PSI domain stimulates higher affinity for (RGD)P2Y₂R on cells, as measured by an increase in the unbinding force. In CHO cells, stably expressing α_IIbβ₃ integrins, virus engagement at the integrin PSI domain, recapitulates physiologic activation of the integrin as indicated by staining with the activation-specific mAB PAC1. The data also show that blocking of the Gα₁₃ protein from binding to the cytoplasmic domain of the β₃ integrin prevents outside-in signaling and infection. We propose that the cis interaction with P2Y₂R provides allosteric resistance to the membrane-normal motion associated with the switchblade model of integrin activation, where the development of tensile force yields physiological integrin activation.

Author List

Bondu V, Wu C, Cao W, Simons PC, Gillette J, Zhu J, Erb L, Zhang XF, Buranda T

Author

Jieqing Zhu PhD Assistant Professor, Associate Investigator in the Biochemistry department at BloodCenter of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
CHO Cells
Cell Adhesion Molecules
Cell Line
Cricetulus
Endothelial Cells
GTP-Binding Protein alpha Subunits, G12-G13
Hantavirus Infections
Humans
Integrin beta Chains
Integrin beta3
Nerve Tissue Proteins
Platelet Glycoprotein GPIIb-IIIa Complex
Protein Binding
Protein Domains
Receptors, Purinergic P2Y2
Semaphorins
Signal Transduction