Search PubMed⌕ Search

Biomedical subjects

J M Kaper

Publications and source records attributed to J M Kaper.

70 records · Page 4Linked to original sources
PubMed1969

Nucleic acid-protein interactions in turnip yellow mosaic virus.

Mild alkaline treatment induces the formation of a specific RNA complex inside turnip yellow mosaic virus. Dialysis at pH 4.5 destroys this complex inside the capsid, but does not destroy complexes that have been removed from their capsids. These facts were interpreted in terms of a breakage and subsequent re-formation of RNA-protein links. In combination with supporting potentiometric data, they suggest histidinyl-phosphate salt links or carboxylate-amino hydrogen bonds, or both, as the principal means of association of turnip yellow mosaic virus protein and RNA.

Centrifugation, Density Gradient↗
PubMedDate not supplied

Identification of a 334-ribonucleotide viral satellite as principal aetiological agent in a tomato necrosis epidemic.

Cucumber mosaic virus (CMV), a widespread and economically important virus of vegetable crops, often contains a satellite RNA, here designated CARNA-5 (for CMV-associated RNA 5). Viral satellites are small nucleic acids that are sequence-unrelated to, but replicatively dependent upon, the viral genome. They essentially are molecular parasites of their helper viruses, and thereby frequently modulate viral symptom expression. Some isolates of CARNA-5 change normally moderate CMV symptoms in tomato into a lethal disease named tomato necrosis; others ameliorate CMV symptoms in tomato and other important crop plants. Here we report on the identification and molecular characterization of a 334-nucleotide necrogenic CARNA-5 isolated from tomato fields in southern Italy, where a massive outbreak of lethal necrosis occurred in the summer of 1988. This is the first time that direct evidence is given for the involvement of a viral satellite in a crop disease of epidemic scale. The possible molecular interrelationships between plant, virus, satellite and other factors that influence the satellite-induced symptom modulation underlying such a catastrophe are discussed.

Fruit↗
PubMedDate not supplied

Cucumber-mosaic-virus-associated RNA-5. XII. Symptom-modulating effect is codetermined by the helper virus satellite replication support function.

In tomato, the disease-modulating effects of a cucumber mosaic virus (CMV) satellite isolate from Belgium, here designated T-CARNA-5 (CARNA-5 = CMV-associated RNA-5), were found to be different depending on the supporting helper virus strain. With two CMV strains, T-CARNA-5 induced lethal necrosis, but with a third strain from Ixora spp. (CMV-Ix), aggravated stunting was observed. However, the primary structure of the T-CARNA-5 contained within virus isolated from tobacco or tomato infected with each of these three CMV strains, conformed to the conserved sequence profile of CARNA-5 isolates which are necrogenic in tomato. Dilution endpoint bioassay of T-CARNA-5 established a direct cause-effect relationship between it and tomato necrosis or stunting, depending on the helper virus. Total nucleic acid extracts taken at different times from tomato plants infected with the above CMV strains and T- or S-CARNA-5 (used as non-necrogenic control) showed viral RNA, ssCARNA-5 and dsCARNA-5 to be present in significant amounts, but in sometimes dissimilar proportions depending on the combination; except in CMV-Ix/S-CARNA-5 infection where neither ss-nor dsCARNA-5 was found. The experiments established that CARNA-5 biological expression studies in CMV-infected tomato have to take into account the helper virus satellite replication support function, which may be a primary codeterminant of quantitative or qualitative differences in the symptom modulation observed.

Base Sequence↗
PubMedDate not supplied

Competition of viral and satellite RNAs of cucumber mosaic virus for replication in vitro by viral RNA-dependent RNA polymerase.

Cucumber mosaic virus (CMV) satellite RNA-induced viral symptom modulation is usually accompanied by a significant reduction of virus accumulation in plant tissue, which has led to the hypothesis that satellite RNA competes with the viral RNAs for replication by the viral replicase and thereby reduces viral RNA synthesis and viral symptoms. In this report, the RNA synthesis of the viral and satellite RNAs of CMV was studied in vitro using an RNA-dependent RNA polymerase (RdRp) purified from CMV-infected plants. Comparison of the kinetics of the CMV RdRp-catalysed RNA synthesis using as templates viral RNAs and satellite RNA, alone or in an appropriate mixture, showed that these RNAs competed with each other for RNA synthesis by the CMV RdRp. Determination of the rates of 32P incorporation into the viral and satellite double-stranded RNA products revealed an apparent replication advantage of the satellite RNA over viral RNAs. The results provide strong support for a previously proposed biochemical mechanism that attributes CMV satellite-induced viral symptom modulation to the replication competition between the satellite and viral RNAs.

Cucumovirus↗
PubMedDate not supplied

Cucumber mosaic virus-associated RNA 5. XIII.--Opposite necrogenicities in tomato of variants with large 5' half insertion/deletion regions.

Two satellite RNA of cucumber mosaic virus (CMV) designated J876-CARNA-5 (for cucumber mosaic virus-associated RNA-5) and D27-CARNA-5 have been molecularly and biologically characterized. J876-CARNA-5 (387 nucleotides (nt)) and D27-CARNA-5 (391 nt) have nearly identical 5' half insertion/deletion regions where 120 nt replace approximately 70 nt of D-CARNA-5 (335 nt), the first variant described and sequenced. J876-CARNA-5 possesses the 15-nt conserved sequence element in its 3' half which is present in all tomato necrogenic variants and induces the same level of necrosis in tomatoes as the prototype necrogenic D-CARNA-5. D27-CARNA-5 lacks the 3' half necrosis-determining element and attenuates the CMV symptoms in tomato. Transcripts of cloned cDNA of J876-CARNA-5 were stably propagated in tomato in the presence of CMV-1. Purified J876-CARNA-5 progeny, inoculated with CMV-1 in a quantitative bioassay, induced tomato necrosis at the same dilution level as the natural satellite. Several computer-generated secondary structures of CMV satellites were examined and the possible correlation of a defined secondary structural element with necrosis induction is discussed.

Base Sequence↗
PubMedDate not supplied

Cucumber mosaic virus from Ixora: infectious RNA transcripts confirm deficiency in satellite support and unusual symptomatology.

A cucumber mosaic virus isolate from an Ixora plant originating in the Philippines and first described 20 years ago (CMV-Ix) by Waterworth and Povish differs from other characterized CMV strains in its anomalous satellite support and in several biological and molecular properties. We describe the preparation of infectious transcripts from cloned complementary DNA and the characterization of progeny virus. The results confirm that CMV-Ix has a more limited host range than most CMV isolates. Virions of CMV-Ix are smaller than those of the control CMV-S, independent of the type of negative stain used. Furthermore, CMV-Ix from transcripts supports the replication of the satellite T-CARNA 5 but not of D-CARNA 5.

Cloning, Molecular↗
PubMedDate not supplied

Effects of electroporation pulse wave on the incorporation of viral RNA into tobacco protoplasts.

The uptake and expression of cucumber mosaic viral (CMV) RNA by tobacco protoplasts was examined using both square wave and exponential wave electroporation pulses. These electropulses, when supplied at sufficient field strength for a critical duration, enabled RNA to be incorporated and expressed in more than 60% of the surviving protoplasts. The results of experiments using these two electroporation wave forms showed significant differences in RNA uptake and expression. The number of viable protoplasts and cells showing expression of RNA was higher over a much broader range of experimental conditions using the square rather than exponential wave generator, even when both machines were optimized for maximal performance. However, at a narrow range of low field strengths the exponential wave pulse generated a higher percentage of transformants than did the square wave pulse. It was shown that after an electroporation pulse from either wave form, there were viable cells which expressed foreign RNA at predictable levels.

Biotechnology↗