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D L Rosen

Publications and source records attributed to D L Rosen.

3 recordsLinked to original sources

Interference between coinoculated viroids.

Experiments were carried out to seek evidence of an interaction between two viroid RNAs introduced to tomato plants in the same inoculum. At the level of symptom expression, the severe isolate of potato spindle tuber viroid (PSTV) dominated the mild isolate. Seventy-five percent of the plants inoculated with a 100-fold excess of the mild isolate developed unattenuated symptoms of severe disease. Other experiments revealed that infectious RNA molecules transcribed from cloned DNA templates containing PSTV sequences reduced the level of hop stunt viroid (HSV) RNA present in nucleic acid extracts of plants which had been inoculated with a mixture of dimeric plus-strand transcripts of these two viroids. Plants inoculated with dual transcripts--containing two copies of PSTV linked to two copies of HSV--developed characteristic symptoms of severe PSTV. Dot hybridization demonstrated that only PSTV replicated to detectable levels in these plants. A likely interpretation of these results is that the HSV portion of the dual transcripts failed to replicate because of interference from PSTV. These results raise questions about how the process of viroid replication is related to symptom expression, and lead to suggested models for the effect of viroid-like RNAs in cells under both normal and pathogenic circumstances.

Plant Diseases

Differential behavior of liposome-introduced specific RNAs in living Drosophila cells.

We have developed a protocol for efficiently introducing macromolecules into Drosophila tissue culture cells using liposomes. By carefully adjusting the fusion parameters, conditions have been established to routinely encapsulate 15-30% of the starting material into liposomes and to introduce 20-30% of the liposome-encapsulated material into the cells during a 30-minute fusion period. Essentially, all of the cells receive material from the liposomes and 10(9) cells can be fused at once. The fusion does not have any measureable effect on cell viability as assayed by trypan blue exclusion, growth rate, and cell morphology. We have utilized this technique to introduce radioactive RNAs into nonradioactive cells, thus enabling the behavior of the introduced RNAs to be followed unambiguously. Liposome-introduced small nuclear RNAs (snRNAs) are stable in the cell for at least 25 hours (approximately two cell generations), with 80% of the radioactivity remaining trichloroacetic acid (TCA) precipitable and the gel electrophoresis pattern remaining essentially unchanged. This is in contrast to liposome-introduced cytoplasmic RNAs, which are only 20% TCA precipitable after the first hour. In the cell, the introduced snRNAs attain a 10-35-fold higher concentration in the nucleus than the cytoplasm. Nuclear accumulation is not seen with Drosophila tRNA or 5S RNA, both of which attain the same nuclear as cytoplasmic RNA concentration.

Animals