1887

Abstract

The p36 and p95 proteins of (CIRV), when expressed in , supported the replication of defective interfering (DI) RNA. Double-label confocal immunofluorescence showed that both proteins localized to mitochondria, independently of each other. DI RNA progeny was localized by hybridization both to mitochondria and to their proximity. Fractionation of cell extracts showed that replicase proteins associated with membranes with a consistent portion of DI RNA. DI RNA transcripts were stabilized more efficiently when co-expressed with both p36 and p95 than with either protein alone. By using the copper-inducible CUP1 promoter, p36 was shown to have an effect on DI RNA stability only above a threshold concentration, suggesting an ‘all-or-none’ behaviour. Conversely, the stabilizing activity of p95 was proportional to protein concentration in the range examined. Similarly, DI RNA replication level was proportional to p95 concentration and depended on a threshold concentration of p36.

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2004-08-01
2019-11-21
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vol. , part 8, pp. 2429-2433

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c-Myc-tagged p36 and HA-tagged p95 support DI RNA replication. Upper panel, representative Northern blot of DI RNA replicated by wild-type (lane 1) and tagged (lane 2) proteins. Lower panel, mean relative accumulation and standard deviation of DI RNA as determined for three independent experiments. Signals were measured with Quantity One software (Bio-Rad).

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