Coat protein sequences of RMV-like strains of barley yellow dwarf virus separate them from other luteoviruses.

Illinois (IL) and Minnesota (MN) RMV-like strains of barley yellow dwarf virus (BYDV) were identified from maize displaying red leaf symptoms by enzyme-linked immunosorbent assay (ELISA) using antiserum against a New York strain (BYDV-RMV-NY). Some IL and MN strains, but not the NY strain, could be detected by ELISA with a monoclonal antibody raised against BYDV-RPV-NY. The region of the viral genome representing the coat protein gene was amplified by polymerase chain reaction, cloned and sequenced. The nucleotide sequences of the BYDV-RMV-IL and BYDV-RMV-MN coat protein genes differed at just five nucleotide positions while the BYDV-RMV-IL and BYDV-RMV-NY differed at 101 of the 591 positions. The predicted amino acid sequences of the coat proteins of RMV-like strains from IL, MN, and NY shared approximately 60% identify with those of the coat proteins of beet western yellows virus, BYDV-RPV-NY, and potato leafroll virus.

Expression strategy of the potato virus X triple gene block.

The mode of expression of the overlapping genes of the triple block positioned internally in potato virus X (PVX) RNA was examined. The results of In vitro translation of synthetic RNA transcripts and natural PVX-specific methylmercuric hydroxide-denatured dsRNAs suggest that the 25K protein is expressed as a single translation product of the 2.1 kb subgenomic (sg) RNA and that both the 12K and 8K proteins are expressed from the same 1.4 kb sgRNA.

In vitro membrane binding of the translation products of the carlavirus 7-kDa protein genes.

Two double-stranded DNA copies of the genes potentially coding for the 7-kDa proteins of potato virus M (PVM) and potato virus S (PVS) were synthesized and cloned into T7 transcription vectors. Cell-free translation of the corresponding monocistronic transcripts yielded in both cases a single protein of approximately 7-8 kDa that contains a highly hydrophobic N-terminal segment. To analyze their membrane-binding potential, both proteins were synthesized in the membrane-enriched Krebs-2 extract. It was found that the smooth membrane fraction was enriched in the carlavirus 7-kDa proteins. The primary and predicted secondary structures of their N-terminal hydrophobic segments suggest that the latter can function as signals for translocation into the rough endoplasmic reticulum.

Expression of RNA transcripts of potato virus X full-length and subgenomic cDNAs.

Translation of synthetic potato virus X (PVX) RNAs was examined in Krebs II ascite cell extracts and rabbit reticulocyte lysates. Either full-length or subgenomic RNAs produced by in vitro transcription of cDNAs cloned in the T7 promoter vectors were used. Full-length PVX RNA-transcript directed the synthesis of a large polypeptide which was indistinguishable from the translation product (165 kDa) of authentic viral genomic RNA on SDS-PAGE. Translation of the full-length and 3'-truncated RNA species was stimulated by addition of m7GpppG to the T7 transcription system. Subgenomic cDNA transcripts corresponding to the 3'-proximal part of PVX RNA were efficiently translated into the PVX coat protein and 2 as yet unidentified small hydrophobic proteins.

[Nucleotide sequence and structural organization of the 3'-terminal region of potato virus M]. / Nukleotidnaia posledovatel'nost' i strukturnaia organizatsiia 3'-kontsevoi oblasti genomnoi RNK M-virusa kartofelia.

The sequence of 2630 3'-terminal nucleotides has been determined for the genomic RNA of potato virus M (PVM), a type member of the carlavirus group. Analysis of this nucleotide sequence revealed five open reading frames coding for proteins of mol. wt. 25, 12, 7, 34 and 11 kDa (in 5'----3' direction). The PVM genome organization has been shown to be basically analogous to that of potexviruses, except that the latter at their 3' end lack the gene encoding 11K protein. Amino acid sequence comparison between the PVM 34K protein and the coat proteins of potexviruses has shown a high extent of homology in the C-terminal amino acids. Homology has also been found between the 25, 12 and 7K proteins encoded by PVM RNA and corresponding potexvirus proteins. All this gave grounds for suggestion that carlaviruses and potexviruses belong to the same subgroup of phytoviruses.

Conserved and variable elements in RNA genomes of potexviruses.

The nucleotide sequences of genomic RNAs and predicted amino acid sequences of two strains of potato virus X and white clover mosaic potexvirus were compared to each other, and the proteins of different plus-RNA-containing plant viruses. The predicted non-virion proteins of potexviruses have direct sequence homology and common structural peculiarities with those of several 'Sindbis-like' plant viruses. The most conserved amino acid sequences were found to be located in the polypeptide encoded by the long 5'-proximal open reading frame (ORF1). The putative polypeptide encoded by the ORF2 starting beyond the ORF1 stop codon is clearly related to the presumptive NTP-binding domain of the ORF1-coded polypeptide. These results suggest possible functions for all of the potexvirus proteins and also indicate that potexviruses have a genome organization which is considerably different from that of other plant viruses.

[Primary structure of the potato virus X genome: the region preceding the capsid protein cistron]. / Izuchenie pervichnoi struktury genoma X-virusa kartofelia: oblast', predshestvuiushchaia tsistronu kapsidnogo belka.

DNA copies of the potato virus X (PVX) RNA corresponding to 2300 nucleotides at the 3'-end have been cloned. The cloned cDNA copies containing the nucleotides 445-1280 from the 3'-end have been sequenced. The 5'-terminal region of the PVX coat protein gene corresponds to residues 445-786 from the 3'-end. The amino acid sequences of two more open reading frames (ORF) have been deduced from the nucleotide sequence. The potential translation products of these ORF's would correspond to the nonstructural viral proteins. We have located the ORF1 within the region of residues 799-1009 preceding the coat protein cistron. The tentative protein is composed of 70 amino acids and has an aminoterminal segment which is markedly hydrophobic. ORF2 in the PVX sequence ends with UAG at nucleotides 942-944 and extends to the 5'-terminus for additional 340 nucleotides. The distant sequence homology exists between a carboxyterminal portion of PVX ORF2 and that of the nonstructural "30 K-proteins" of the plant tobamoviruses.

[Salmonella typhimurium as a test system for detecting the mutagenic activity of environmental pollutants. I. The mutagenic action of heavy metal salts in in vivo and in vitro systems without metabolic activation]. / Salmonella typhimurium--test-sistema dlia vyiavleniia mutagennoi aktivnosti zagriaznitelei okruzhaiushchei sredy. Soobshchenie I. Mutagennoe deistvie solei tiazhelykh metallov v sistemakh in vivo i in vitro bez metabolicheskoi aktivatsii.

The work presents the data on mutagenic effects of heavy metal salts (Zn and Cd) on Salmonella typhimurium test strains using mutagenicity test in vitro without metabolic activation and host-mediated assay. The techniques used enabled to determine also the types of mutations arising from the exposure to ZnCl2 and CdCl2.

[Salmonella typhimurium--test system for detecting the mutagenic activity of environmental pollutants. II. Mutagenic effect of heavy metal salts in an in vitro system with metabolic activation]. / Salmonella typhimurium--test-sistema dlia vyiavleniia mutagennoi aktivnosti zagriaznitelei okruzhaiushchei sredy. Soobshchenie II. Mutagennyi éffekt solei tiazhelykh metallov v sisteme in vitro s metabolicheskoi aktivatsiei.

Mutagenic effect of zinc chloride on Salmonella typhimurium strain was detected using in vitro metabolic activation system. Cadmium chloride showed no significant mutagenic activity in the same system. It is recommended to use both in vitro and in vivo metabolic activation systems in mutagenicity testing of chemicals.