RNA polymerase activity and poly(A) synthesizing activity in defective T particles of vesicular stomatitis virus

Reichmann, M. E.; Villarreal, Louis Perez; Kohne, David E.; Lesnaw, Judith A.; Holland, John J.

doi:10.1016/0042-6822(74)90158-5

Cited by 38 publications

(28 citation statements)

References 20 publications

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“…The rate of protein synthesis in response to added mRNA was usually linear for 30-45 min and then gradually declined. In contrast, under coupled conditions there was an initial lag phase of [15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30] min, followed by linear rates of protein synthesis for up to 2.5 hr. These results were repeatedly observed in the wheat embryo extract (Fig.…”

Section: Translationmentioning

confidence: 99%

“…Annealing reactions were carried out in sealed glass capillaries at 70°for 20 hr as described elsewhere (16) (2) and with (3) addition of VSV in vitro mRNA, proteins synthesized in the rabbit reticulocyte extract without (4) and with (5) VSV in vitro mRNA, and proteins synthesized in the wheat embryo extract without (6) and with (7) VSV in vitro mRNA. The autoradiograms shown in slots 1-7 were exposed for 2 days.…”

Section: )mentioning

confidence: 99%

“…The Indiana serotype of VSV was grown in BHK 21 (baby hamster kidney) cells and purified by sucrose gradient centrifugation and subsequent potassium tartrate gradient centrifugation, as described elsewhere (16 (17) and purified by sucrose gradient centrifugation (18).…”

Section: )mentioning

confidence: 99%

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Coupled in vitro transcription and translation of vesicular stomatitis virus messenger RNA.

Breindl¹,

Holland²

1975

Proc. Natl. Acad. Sci. U.S.A.

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proteins (3-5) are the glycoprotein (G), the matrix protein (M), the nucleoprotein (N) and two minor proteins (L and NS). VSV also has a virion-associated RNA polymerase (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) which can synthesize RNA complementary to the viral genome in vitro (6, 7). The in vitro RNA products have molecular weights ranging from 0.2 to 1 X 106 (8) and are polyadenylylated (9,

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Section: Translationmentioning

confidence: 99%

Section: )mentioning

confidence: 99%

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Coupled in vitro transcription and translation of vesicular stomatitis virus messenger RNA.

Breindl¹,

Holland²

1975

Proc. Natl. Acad. Sci. U.S.A.

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“…

The function of these VSV-specific small RNAs is unknown.

…”

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confidence: 99%

“…More recently, small RNAs that are synthesized in vtro by purified VSV DI particles have been characterized and sequenced (3)(4)(5)(6)(7)(8). These small RNAs resemble the "leader" sequence described by Colonno and Banerjee (9)(10)(11) in that they contain 46-48 nucleotides with a (p)ppAp at the 5' end and they lack poly(A) at the 3' end.…”

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confidence: 99%

Synthesis of a small RNA in cells coinfected by standard and defective interfering particles of vesicular stomatitis virus.

Rao¹,

Huang²

1979

Proc. Natl. Acad. Sci. U.S.A.

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(3)(4)(5)(6)(7)(8). These small RNAs resemble the "leader" sequence described by Colonno and Banerjee (9-11) in that they contain 46-48 nucleotides with a (p)ppAp at the 5' end and they lack poly(A) at the 3' end.The function of these VSV-specific small RNAs is unknown. To study their function, conditions were sought for reproducibly detecting their synthesis in infected cells. Neither standard infectious VSV nor DI particles of VSV alone led to the synthesis of any small virus-specific RNA. Only when cells were coinfected with standard and DI particles was a small RNA detected. The synthesis of this small RNA correlated directly with the replication of the genome of DI particles. Both RNA species were absent from coinfected cells when cycloheximide was present or when DI particles were irradiated with UV light prior to infection. These data can be interpreted in terms of interference with the growth of standard VSV by DI particles. MATERIALS AND METHODSCells and Viruses. Suspension cultures of baby hamster kidney (BHK) cells, originally obtained from Amiya Banerjee, were grown in Joklik's modified minimal essential medium supplemented with 5% fetal calf serum. Cloned and sucrose gradient-purified standard infectious particles of VSV of the Indiana serotype (San Juan strain) and two different DI particles derived from the same San Juan standard VSV were used (12, 13). The DI-T particle has been characterized in detail (14-18). It is one-third the size of standard VSV and contains sequences from the 5' end of the genome. DI0.52 particles were selected from cloned VSV by multiple undiluted passages in mouse L cells and then subsequent growth in BHK cells. DI0.52 particles are twice the size of DI-T particles and also map at the 5' end of the VSV genome (unpublished observations). Both DI particles were sucrose gradient-purified and their multiplicities were determined as described (19).Labeling of Infected Cells. BHK cells were infected at a multiplicity of 20 with either standard VSV or DI particles alone or were coinfected with equal amounts of standard VSV or DI particles at a total multiplicity of 40. Detailed procedures for infection and labeling with 32P in the presence of actinomycin D have been published (1).Preparation and Analysis of RNA. RNAs from VSV particles or from infected cells were extracted as described (1). To analyze total cytoplasmic RNA, ethanol-precipitated RNA was resuspended in 5 ,gl of deionized H20 and mixed with an equal volume of double-strength electrophoresis buffer [6 M urea, the dyes xylene cyanol (XC) and bromphenol blue, and 0.01% sucrose]. To denature the large RNA species fully, they were first treated with 90% dimethyl sulfoxide and heated at 50'C for 30 min. This treatment was not necessary for small RNA. For the separation of virion RNA and mRNA (large RNAs), 1.5% agarose/6 M urea, pH 3.8 was used (1,20). Electrophoresis was at 100 V at 40C until the XC was 5 cm from the bottom of the gel. Then, the 1.5% agarose gel was dried and exposed to x-ray film as described (1).Fo...

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Defective Interfering RNA Viruses and the Host-Cell Response

Holland

Kennedy

Semler

et al. 1980

Comprehensive Virology

124

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This chapter will emphasize recently derived knowledge concerning the nature of defective interfering (DI) particles of RNA animal viruses, their biological origins and functions, and their involvement in long-term persistent infections. We will not attempt to review all of the DI literature, and we will confine ourselves to DI particles of RNA viruses. The previous review by Huang and Baltimore (1977) amply documents the occurrence and behavior of DI particles in a wide variety of DNA and RNA viruses and discusses their biological effects, and a very thorough recent review of rhabdovirus DI particles by Reichmann and Schnitzlein (1978) provides excellent in-depth coverage of many areas not covered by the present chapter as well as some alternate viewpoints of areas which are considered here. We will omit DNA virus DI particles from extensive consideration because of space limitations and because they are generally less well characterized at present.

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RNA polymerase activity and poly(A) synthesizing activity in defective T particles of vesicular stomatitis virus

Cited by 38 publications

References 20 publications

Coupled in vitro transcription and translation of vesicular stomatitis virus messenger RNA.

Coupled in vitro transcription and translation of vesicular stomatitis virus messenger RNA.

Synthesis of a small RNA in cells coinfected by standard and defective interfering particles of vesicular stomatitis virus.

Defective Interfering RNA Viruses and the Host-Cell Response

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