The Ribonucleic Acids of the Infective and Interfering Components of Vesicular Stomatitis Virus

Abstract

SUMMARYThe RNA-containing components of vesicular stomatitis virus were labelled with asp by growing the virus in a phosphate-deficient medium containing a2PO 4 and Actinomycin-D. Centrifugation of the virus suspensions in a sucrose gradient gave four radioactive fractions. The most rapidly sedimenting fraction contained the infective component of the virus, whereas most of the interfering activity was in the second fraction, which contained 'caps', filaments and rosette-like structures. The interfering activity was associated with the 'cap', which could be separated from the filaments and rosettes by centrifuging in a potassium tartrate gradient.The viral RNA sedimented predominantly in the 36 to 4o S position in sucrose gradients prepared in o.I M-acetate buffer but frequently exhibited a more heterogeneous profile. The percentage distribution of 32p in the four nucleotides was A = 29"3, C = 2i.i, G = 2o'9 and U = 28"7. Ribonucleic acid from the interfering component invariably gave a sharp profile with a sedimentation coefficient of I8 to 2oS and the percentage distribution of 32p was A = 27"I, C = 2I'9, G = 2o'3 and U --30"7. Both nucleic acids were hydrolysed to low molecular weight materials by ribonuclease o.m /zg./ml.

Characterization of Snap-back RNAs in Vesicular Stomatitis Defective Interfering Virus Particles

“…passages following cloning (Reichmann, Pringle & Follett, 197I ;Stampfer, Baltimore & Huang, 1971 ;Perrault & Holland, i972a;Holland, Villarreal & Breindl, I976a). These shorter truncated or T particles contain deleted forms of the genome or B RNA (Huang & Wagner, 1966;Brown et al 1967;Schaffer, Hackett & Soergel, 1968;Roy & Bishop, 1972) which specifically inhibit the replication of the latter (Huang & Manders,I972;Perrault & Holland,I972b).…”

Comprehensive Virology

“…Huang and Wagner (l966b) extracted the RNA frorn infective VS B virions and estimated its molecular weight at 3.1-4.0 x 10 6 daltons based on a sedimentation coefficient of 42 S. Similar values, ranging from 38 S to 43 S, have been obtained subsequently by many investigators (Brown et al, 1967c). Comparable values have also been obtained by polyacrylamide gel electrophoresis; by this method the molecular weight of VS B-virion RNA was estimated to be 4.4 X 10 6 daltons at the upper extreme (Bishop and Roy, 1971a) and 3.2 x 10 6 daltons at the lower extreme (Kiley and Wagner, 1972).…”

“…Hackett (1964) demonstrated a possible morphological basis for this auto interference phenomenon which was subsequently proven by definitive identification and purification of a truncated (T) particle in undiluted-passage preparations of VS virus Huang and Wagner, 1966a;Brown et al, 1967c). When such preparations are examined by PTA negative-staining electron microscopy, the predominant virions are about one-third the length (about 65 nm) of Fig.…”

Replication of Vesicular Stomatitis Virus: the Effect of Purified Interfering Component

“…Under these conditions, an excess of smaller non-infectious particles is synthesized which has been shown to inhibit the synthesis of infectious virus (Crick,Cartwright & Brown,I966;Huang & Wagner, I966; Hackett,Schaffer & Madin,t967). The purified interfering component contains an RNA which is smaller than the virus RNA (Huang & Wagner, I966;Brown et al 1967) but has the same polypeptides (Kang & Prevec,I969;Wagner,Schnaitman & Snyder,I969) and is antigenicalty indistinguishable from the virus (Huang, Greenawalt & Wagner, 1966). Huang & Baltimore (I97O) have postulated that interfering component arises from a deletion mutant in the parent virus.…”