Enhanced Fusion Pore Expansion Mediated by the Trans-Acting Endodomain of the Reovirus FAST Proteins

The reovirus fusion-associated small transmembrane (FAST) proteins are virus-encoded membrane fusion proteins that function as dedicated cell–cell fusogens. The topology of these small, single-pass membrane proteins orients the majority of the protein on the distal side of the membrane (i.e., inside the cell). We now show that ectopic expression of the endodomains of the p10, p14, and p15 FAST proteins enhances syncytiogenesis induced by the full-length FAST proteins, both homotypically and heterotypically. Results further indicate that the 68-residue cytoplasmic endodomain of the p14 FAST protein (1) is endogenously generated from full-length p14 protein expressed in virus-infected or transfected cells; (2) enhances syncytiogenesis subsequent to stable pore formation; (3) increases the syncytiogenic activity of heterologous fusion proteins, including the differentiation-dependent fusion of murine myoblasts; (4) exerts its enhancing activity from the cytosol, independent of direct interactions with either the fusogen or the membranes being fused; and (5) contains several regions with protein–protein interaction motifs that influence enhancing activity. We propose that the unique evolution of the FAST proteins as virus-encoded cellular fusogens has allowed them to generate a trans-acting, soluble endodomain peptide to harness a cellular pathway or process involved in the poorly understood process that facilitates the transition from microfusion pores to macrofusion and syncytiogenesis

A Virus-Encoded Cell–Cell Fusion Machine Dependent on Surrogate Adhesins

The reovirus fusion-associated small transmembrane (FAST) proteins function as virus-encoded cellular fusogens, mediating efficient cell–cell rather than virus–cell membrane fusion. With ectodomains of only ∼20–40 residues, it is unclear how such diminutive viral fusion proteins mediate the initial stages (i.e. membrane contact and close membrane apposition) of the fusion reaction that precede actual membrane merger. We now show that the FAST proteins lack specific receptor-binding activity, and in their natural biological context of promoting cell–cell fusion, rely on cadherins to promote close membrane apposition. The FAST proteins, however, are not specifically reliant on cadherin engagement to mediate membrane apposition as indicated by their ability to efficiently utilize other adhesins in the fusion reaction. Results further indicate that surrogate adhesion proteins that bridge membranes as close as 13 nm apart enhance FAST protein-induced cell–cell fusion, but active actin remodelling is required for maximal fusion activity. The FAST proteins are the first example of membrane fusion proteins that have specifically evolved to function as opportunistic fusogens, designed to exploit and convert naturally occurring adhesion sites into fusion sites. The capacity of surrogate, non-cognate adhesins and active actin remodelling to enhance the cell–cell fusion activity of the FAST proteins are features perfectly suited to the structural and functional evolution of these fusogens as the minimal fusion component of a virus-encoded cellular fusion machine. These results also provide a basis for reconciling the rudimentary structure of the FAST proteins with their capacity to fuse cellular membranes

Pressure distribution in a reservoir affected by capillarity and hydrodynamic drive: Griffin Field, North West Shelf, Australia

The effects of capillarity in a multilayered reservoir with flow in the aquifer beneath have characteristic signatures on pressure-elevation plots. Such signatures are observed for the Griffin and Scindian/Chinook fields of the Carnarvon Basin North West Shelf of Australia. The Griffin and Scindian/Chinook fields have a highly permeable lower part to the reservoir, a less permeable upper part, and a low permeability top seal. In the Griffin Field there is a systematic tilt of the free-water level in the direction of groundwater flow. Where the oil-water contact occurs in the less permeable part of the reservoir, it lies above the free-water level due to capillarity. In addition to these observable hydrodynamic and capillary effects on hydrocarbon distribution, the multi-well pressure analysis shows that the gas-oil contacts in the Scindian/Chinook fields occur at different elevations. For both the Griffin and Scindian/Chinook fields the oil pressure gradients from each well are non-coincident despite continuous oil saturation, and the difference is not attributable to data error. Furthermore, the shift in oil pressure gradient has a geographical pattern seemingly linked to the hydrodynamics of the aquifer. The simplest explanation for all the observed pressure trends is an oil leg that is still in the process of equilibrating with the prevailing hydrodynamic regime