Dissection of Vesicular Stomatitis Virus into the Infective Ribonucleoprotein and Immunizing Components

Abstract

SUMMARYTreatment of the infective component of vesicular stomatitis virus with Nonidet P4o produces an infective skeleton-like structure which has the shape and approximate size of the intact virus particle. The infectivity of the skeleton is enhanced I oo to I ooo fold by mixing with DEAE-dextran. The skeleton lacks the outer envelope and fringe structure and in consequence does not produce neutralizing antibodies in guinea pigs. The density of the skeleton is 1.22 g./ml, in potassium tartrate gradients compared with x "t4 g./ml, for the virus. Sodium deoxycholate removes protein from the skeleton and releases the filamentous ribonucleoprotein in an infective form. As with the skeleton, the infectivity of the ribonucleoprotein is enhanced by DEAEdextran. Ribonuclease has no effect on the ribonucleoprotein but trypsin destroys its infectivity. The ribonucleoprotein has a density of ~-22 g./ml, in tartrate gradients, sediments at about ~40 s in sucrose gradients and does not produce neutralizing antibodies in guinea-pigs.

Intergenotypic Replacement of Lyssavirus Matrix Proteins Demonstrates the Role of Lyssavirus M Proteins in Intracellular Virus Accumulation

“…Since the late domain PPEY motif in the M proteins of lyssaviruses is conserved, we did not expect a typical late domain-defective phenotype with virus accumulations at the surface of the infected cells. Defects in the assembly pathway could also lead to so-called skeleton-RNPs, which are condensed, M containing nonenveloped subviral structures that are observed after detergent treatment of enveloped VSV and RABV virions (5,32). Moreover, accumulation of noncondensed RNPs as a result of defects in RNP recruitment by M may occur.…”

Conformation of the helical nucleocapsids of paramyxoviruses and vesicular stomatitis virus: reversible coiling and uncoiling induced by changes in salt concentration.

“…A striking demonstration of the effect of the environment on nucleocapsid conformation is the fact that in the VSV virion the nucleocapsid is wound into a coil with a diameter about twice that seen when it is released from virions and coiled in the presence of salt (13,(15)(16)(17) (Fig. 3b).…”

“…Two other proteins (L and P) are associated with the nucleocapsid and thought to be involved in the viral RNA transcriptase activity (8)(9)(10)(11)(12). The nucleocapsids of rhabdoviruses-e.g., vesicular stomatitis (VSV) and rabies viruses-also consist of a single-stranded RNA complexed with a single major structural protein (N) and two associated proteins (NS and L) (13)(14)(15)(16)(17)(18). In the VSV virion, the nucleocapsid forms a coil t40 nm in diameter but, when separated from the viral membrane, it is often found in helices of considerably smaller diameter (13)(14)(15)(16)(17).…”

“…When released from virions, the nucleocapsids of rhabdoviruses form helices -20 nm in diameter (13,(14)(15)(16)(17). The effect of salt concentration on VSV nucleocapsids was examined and, as with paramyxoviruses, the VSV nucleocapsid was found to unwind reversibly at low salt concentration.…”

Quantification of the Antiviral Effect of Interferon by Immunoassay of Vesicular Stomatitis Virus Proteins

“…Comparison of several detergents suggested a correlation between an increase in binding to the PVC and the effectiveness of the detergent in dissociating the proteins of VSV. Thus, deoxycholate which causes release of both of the G and M proteins of VSV and a disruption of the viral nucleocapsid (Cartwright et al, 1970a, b) was significantly more effective than NP40, which is known to release selectively only the G protein (Cartwright et al, 1970a, b) (Fig. 1 a).…”