The Fusion Loops of the Initial Prefusion Conformation of Herpes Simplex Virus 1 Fusion Protein Point Toward the Membrane

All enveloped viruses, including herpesviruses, must fuse their envelope with the host membrane to deliver their genomes into target cells, making this essential step subject to interference by antibodies and drugs. Viral fusion is mediated by a viral surface protein that transits from an initial prefusion conformation to a final postfusion conformation. Strikingly, the prefusion conformation of the herpesvirus fusion protein, gB, is poorly understood. Herpes simplex virus (HSV), a model system for herpesviruses, causes diseases ranging from mild skin lesions to serious encephalitis and neonatal infections. Using cryo-electron tomography and subtomogram averaging, we have characterized the structure of the prefusion conformation and fusion intermediates of HSV-1 gB. To this end, we have set up a system that generates microvesicles displaying full-length gB on their envelope. We confirmed proper folding of gB by nondenaturing electrophoresis-Western blotting with a panel of monoclonal antibodies (MAbs) covering all gB domains. To elucidate the arrangement of gB domains, we labeled them by using (i) mutagenesis to insert fluorescent proteins at specific positions, (ii) coexpression of gB with Fabs for a neutralizing MAb with known binding sites, and (iii) incubation of gB with an antibody directed against the fusion loops. Our results show that gB starts in a compact prefusion conformation with the fusion loops pointing toward the viral membrane and suggest, for the first time, a model for gB’s conformational rearrangements during fusion. These experiments further illustrate how neutralizing antibodies can interfere with the essential gB structural transitions that mediate viral entry and therefore infectivity

Fedosov Quantization of Lagrange-Finsler and Hamilton-Cartan Spaces and Einstein Gravity Lifts on (Co) Tangent Bundles

We provide a method of converting Lagrange and Finsler spaces and their Legendre transforms to Hamilton and Cartan spaces into almost Kaehler structures on tangent and cotangent bundles. In particular cases, the Hamilton spaces contain nonholonomic lifts of (pseudo) Riemannian / Einstein metrics on effective phase spaces. This allows us to define the corresponding Fedosov operators and develop deformation quantization schemes for nonlinear mechanical and gravity models on Lagrange- and Hamilton-Fedosov manifolds.Comment: latex2e, 11pt, 35 pages, v3, accepted to J. Math. Phys. (2009

Teleparallel Lagrange Geometry and a Unified Field Theory

In this paper, we construct a field theory unifying gravity and electromagnetism in the context of Extended Absolute Parallelism (EAP-) geometry. This geometry combines, within its structure, the geometric richness of the tangent bundle and the mathematical simplicity of Absolute Parallelism (AP-) geometry. The constructed field theory is a generalization of the Generalized Field Theory (GFT) formulated by Mikhail and Wanas. The theory obtained is purely geometric. The horizontal (resp. vertical) field equations are derived by applying the Euler-Lagrange equations to an appropriate horizontal (resp. vertical) scalar Lagrangian. The symmetric part of the resulting horizontal (resp. vertical) field equations gives rise to a generalized form of Einstein's field equations in which the horizontal (resp. vertical) energy-momentum tensor is purely geometric. The skew-symmetric part of the resulting horizontal (resp. vertical) field equations gives rise to a generalized form of Maxwell equations in which the electromagnetic field is purely geometric. Some interesting special cases, which reveal the role of the nonlinear connection in the obtained field equations, are examined. Finally, the condition under which our constructed field equations reduce to the GFT is explicitly established.Comment: Latex file, 33 page

Herpesvirus Glycoproteins Undergo Multiple Antigenic Changes before Membrane Fusion

Herpesvirus entry is a complicated process involving multiple virion glycoproteins and culminating in membrane fusion. Glycoprotein conformation changes are likely to play key roles. Studies of recombinant glycoproteins have revealed some structural features of the virion fusion machinery. However, how the virion glycoproteins change during infection remains unclear. Here using conformation-specific monoclonal antibodies we show in situ that each component of the Murid Herpesvirus-4 (MuHV-4) entry machinery—gB, gH/gL and gp150—changes in antigenicity before tegument protein release begins. Further changes then occurred upon actual membrane fusion. Thus virions revealed their final fusogenic form only in late endosomes. The substantial antigenic differences between this form and that of extracellular virions suggested that antibodies have only a limited opportunity to block virion membrane fusion

The herpevac trial for women: Sequence analysis of glycoproteins from viruses obtained from infected subjects

<div><p>The Herpevac Trial for Women revealed that three dose HSV-2 gD vaccine was 58% protective against culture-positive HSV-1 genital disease, but it was not protective against HSV-2 infection or disease. To determine whether vaccine-induced immune responses had selected for a particular gD sequence in strains infecting vaccine recipients compared with viruses infecting control subjects, genetic sequencing studies were carried out on viruses isolated from subjects infected with HSV-1 or HSV-2. We identified naturally occurring variants among the gD sequences obtained from 83 infected subjects. Unique or low frequency amino acid substitutions in the ectodomain of gD were found in 6 of 39 HSV-1-infected subjects and in 7 of 44 HSV-2-infected subjects. However, no consistent amino acid change was identified in isolates from gD-2 vaccine recipients compared with infected placebo recipients. gC and gE surround and partially shield gD from neutralizing antibody, and gB also participates closely in the viral entry process. Therefore, these genes were sequenced from a number of isolates to assess whether sequence variation may alter protein conformation and influence the virus strain’s capacity to be neutralized by vaccine-induced antibody. gC and gE genes sequenced from HSV-1-infected subjects showed more variability than their HSV-2 counterparts. The gB sequences of HSV-1 oral isolates resembled each other more than they did gB sequences rom genital isolates. Overall, however, comparison of glycoprotein sequences of viral isolates obtained from infected subjects did not reveal any singular selective pressure on the viral cell attachment protein or surrounding glycoproteins due to administration of gD-2 vaccine.</p></div