Temperature-sensitive mutants of vesicular stomatitis virus

Cormack, D. V.; Holloway, A. F.; Wong, Patty; Cairns, J. E.

doi:10.1016/0042-6822(71)90205-4

Cited by 13 publications

(4 citation statements)

References 4 publications

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“…Therefore, mutants in complementation group I represent either a mutation in the transcriptase gene, or a defect of template which appears at nonpermissive temperature; one region of the ts 5 template may not be "accessible" to the polymerase at 39.2 C. This phenomenon could be related to the one observed by S. V. Emerson and R. R. Wagner (in press) with T particles. In good agreement with our data, Printz and Wagner (25) and, recently, Wunner and Pringle (34) did not detect any viral protein synthesis at the restrictive temperature with ts mutants of group I. Transcriptase-defective VS virions have also been identified by several authors (6,30) although it has not yet been proved that these mutants belong to the same complementation group.…”

Section: Discussionsupporting

confidence: 92%

Study of the Transcription and the Replication of Vesicular Stomatitis Virus by Using Temperature-Sensitive Mutants

Printz-Ane

Combard

Martinet

1972

J Virol

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The viral ribonucleic acids (RNA species) synthesized in HeLa cells infected with temperature-sensitive (ts) mutants and with the wild type of vesicular stomatitis virus at permissive (30 C) and nonpermissive (39.2 C) temperatures were compared by sucrose gradient centrifugation. Two ts mutants (ts 5 and ts 100) representing two separate complementation groups (respectively, groups I and IV), each concerned with viral RNA synthesis, were chosen. Mutant ts 5 failed to synthesize any RNA at 39.2 C. Under the same conditions, mutant ts 100 showed a low, but easily detectable, synthesis of RNA without characteristic peaks. The in vitro transcriptase activity was tested with mutants ts 5 and ts 100 at 39.2 C: normal activity, compared with wild-type virus, was detected with purified ts 100, but no activity was detected with purified ts 5. From all our data we conclude that mutant ts 5 is defective in transcription. The defect could be in the structural transcriptase enzyme or at the level of template for transcription. Results with mutant ts 100 were not so clear-cut. However, we suggest that this mutant is concerned with some aspect of transcription in vivo. In addition, our results lead us to postulate some linkage between transcription and replication.

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Section: Discussionsupporting

confidence: 92%

Study of the Transcription and the Replication of Vesicular Stomatitis Virus by Using Temperature-Sensitive Mutants

Printz-Ane

Combard

Martinet

1972

J Virol

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“…Mutants of complementation groups I and IV (7,12,19) are classified as RNA-owing to marked reduction of' RNA synthesis in vivo at restrictive temperatures (17,20,26). Definitive evidence has also been provided for defective transcription at the restrictive temperature for certain group I mutants (6,8,21,22). There appears to be some variability among the mutants in complementation group IV, the RNAphenotype of which may or may not be related to defective transcription (6,21,22).…”

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confidence: 99%

Location of the Transcription Defect in Group I Temperature-Sensitive Mutants of Vesicular Stomatitis Virus

Hunt

Wagner

1974

J Virol

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The ribonucleoprotein-dependent RNA transcriptase in vesicular stomatitis B virions of four temperature-sensitive (ts) mutants belonging to complementation group I was analyzed in vitro at permissive (31 C) and restrictive (39 C) temperatures. The RNA-synthesizing activity of all four ts mutants was more labile at 39 C than was the transcriptive activity of wild-type (wt) virions. In order to locate the temperature-sensitive transcription defect in the mutants, wt and ts mutant virions were fractionated by Triton X-100-high salt solubilizer into a sedimentable ribonucleoprotein template and a nonsedimentable enzyme fraction, each of which alone had little or no transcriptive activity. The templateand enzyme-containing fractions of wt virions were then tested for their capacity to restore transcriptive activity at 39 C to corresponding template and enzyme preparations of ts mutant virions. Recombination of' wt template and ts enzymes resulted in no significant restoration of capacity to synthesize RNA at restrictive temperature. In contrast, transcriptive function at 39 C was reconstituted by recombining the wt enzyme with the template component of ts mutants. It appears, therefore, that the enzyme, rather than the template, is the temperature-sensitive component of the transcription complex of group I vesicular stomatitis virus mutants.

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“…F. Howatson,personal communication) and classified into groups by complementation tests. The physiological properties of these mutants are being investigated in detail, either to provide a picture of the organisation of the genome of VSV or to provide experimental tools for investigation of virus multiplication (Lafay, 1969, I97I;Lafay & Berkaloff, I969;Deutsch, ~97o;Martinet & Printz-Ane, I97o;Cormack et al 197I;Deutsch & Berkaloff, I971 ;Pringle & Duncan, I97I ;Printz & Wagner, I971 ;Reichman, Pringle & Follett, I97I ;Wunner & Pringle, I972;Szilfigyi & Pringle, I972;Wong, Holloway & Cormack, I972;Zfivada, I972). Consequently, it is important to establish the precise genetic homologies of these mutants to facilitate comparison of results obtained in different laboratories.…”

Section: Introductionmentioning

confidence: 99%