Anomalous electrophoretic behavior of the major potato virus X RNA translation product

Karasev, Alexander V.; Miroshnichenko, N.A.; Rozanov, Mikhail

doi:10.1016/0166-0934(89)90136-5

Cited by 6 publications

(4 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The experimental molecular mass was 200 kDa (Fig. 4B, lane 4), but the ORF 1 product is known to migrate anomalously in Tris-glycine-SDS-polyacrylamide gels (12). Four viral proteins (24,12,7, and 25 kDa) were detected along with minor protein products (18 to 22 kDa) when total protein extracts from infected leaf tissue were incubated with a mixture of peptide antisera and coat protein antiserum (Fig.…”

Section: Vims D=nmentioning

confidence: 96%

“…1.Despite the detection of seven viral RNA species, only two virus particle sizes have been detected by Grama and Mashkovsky (7). The larger-particle population contained genomic RNA (6.4 kb), and the other population contained a 0.9-kb RNA.The coding properties of PVX genomic RNA and the 0.9-kb RNA were examined in in vitro translation systems (7,12). Two major translation products, 165 and 25 kDa, corresponding to ORFs 1 and 5, respectively, were synthesized in rabbit reticulocyte lysate and wheat germ extracts t Present address:…”

mentioning

confidence: 99%

mentioning

confidence: 99%

“…The coding properties of PVX genomic RNA and the 0.9-kb RNA were examined in in vitro translation systems (7,12). Two major translation products, 165 and 25 kDa, corresponding to ORFs 1 and 5, respectively, were synthesized in rabbit reticulocyte lysate and wheat germ extracts t Present address: Department of Plant Pathology, Johnson Hall, Washington State University, Pullman, WA 99164-6430. from 6.4-and 0.9-kb viral RNAs extracted from purified virus preparations (7,12). Recently, synthetic RNA transcripts were produced from truncated cDNA clones spanning different regions of PVX genomic RNA.…”

mentioning

confidence: 99%

See 3 more Smart Citations

Examination of potato virus X proteins synthesized in infected tobacco plants

Price

1992

J Virol

View full text Add to dashboard Cite

Nucleotide sequence analysis of potato virus X (PVX) genomic RNA predicts five open reading frames (ORFs). Previous analysis of total RNAs from PVX-infected leaf tissue suggested that six subgenomic RNAs are synthesized during infection. However, the proteins encoded by the genomic RNA, the subgenomic RNAs, or the predicted ORFs have not been identified in vivo. To characterize the coding properties of the viral RNA, particularly to determine whether the five predicted ORFs function in vivo, total protein extracts prepared from PVX-infected leaf tissue were analyzed by using antibodies raised against virus-specific synthetic peptides and against the virus capsid protein. Dot blot analyses showed that these antibodies reacted to PVX-infected extracts, indicating in vivo expression of the five predicted ORFs. In addition, Western blot (immunoblot) analysis of the extracts showed that ORF 1, 2, 3, and 4 peptide antisera and coat protein antiserum detect predominantly a single protein.Potato virus X (PVX) is the type member of the potexviruses (2). The virus contains a single-stranded, positivesense RNA about 6,400 nucleotides in length (2). Computer analysis of the nucleotide sequence of PVX genomic RNA predicts five open reading frames (ORFs) encoding proteins of 165 (ORF 1), 24 (ORF 2), 12 (ORF 3), 7 (ORF 4), and 25 (ORF 5) kDa ( Fig. 1) (8). Other potexviruses, including white clover mosaic virus, papaya mosaic virus, and narcissus mosaic virus, share a similar genomic organization (6,20,24).Despite the large coding capacity of PVX genomic RNA, in vitro translation studies generally yielded high-molecularmass proteins, 145 and 180 kDa (19, 23). These proteins do not account for the total coding capacity of the viral RNA. Hybrid arrest translation studies indicated that the 180and 145-kDa proteins were derived from the 5' end of the genome (23). Dolja et al. (4) demonstrated that virus-related singleand double-stranded RNAs could be isolated from infected plants. In addition to full-length genomic RNA, they observed six 3' coterminal poly(A)-containing subgenomic RNA species. Two RNA species, 2.1 and 0.9 kb in length, were present at higher concentrations than the other four, which were 3.6, 3.0, 1.8, and 1.4 kb in length. Doublestranded RNA counterparts of most of these RNAs were also observed. Two subgenomic RNAs, 3.6 and 3.0 kb, do not have apparent corresponding ORFs. ORFs corresponding to these RNAs are denoted by question marks in Fig. 1.Despite the detection of seven viral RNA species, only two virus particle sizes have been detected by Grama and Mashkovsky (7). The larger-particle population contained genomic RNA (6.4 kb), and the other population contained a 0.9-kb RNA.The coding properties of PVX genomic RNA and the 0.9-kb RNA were examined in in vitro translation systems (7,12). Two major translation products, 165 and 25 kDa, corresponding to ORFs 1 and 5, respectively, were synthesized in rabbit reticulocyte lysate and wheat germ extracts t Present address:

show abstract

Section: Vims D=nmentioning

confidence: 96%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Examination of potato virus X proteins synthesized in infected tobacco plants

Price

1992

J Virol

View full text Add to dashboard Cite

show abstract

The Movement Protein-Triggered in Situ Conversion of Potato Virus X Virion RNA from a Nontranslatable into a Translatable Form

et al. 2000

View full text Add to dashboard Cite

Plant virus-encoded movement protein(s) (MP), and for many viruses the coat protein (CP), is required to mediate viral spread between plant cells via plasmodesmata (PD). Most probably, the genomic RNA of potexviruses moves through PD as assembled virions and/or as ribonucleoprotein complexes containing the CP and 25-kDa MP. Here we report that encapsidated potato virus X (PVX) virion RNA, which is nontranslatable in a cell-free protein synthesizing system, can be converted into a fully translatable form after interaction of intact PVX particles with the PVX 25-kDa MP. The 25-kDa MP molecules bind selectively to only one extremity of the viral particle (that presumably contains the 5' end of the genomic RNA). The process of complex formation is ATP-independent; i.e., the ATPase activity of the 25-kDa MP is not involved in the binding of the MP to PVX virion.

show abstract

Translational Activation of Encapsidated Potato Virus X RNA by Coat Protein Phosphorylation

et al. 2001

View full text Add to dashboard Cite

Previously we showed that encapsidated potato virus X (PVX) RNA is nontranslatable in vitro, but can be converted into a translatable form after binding to PVX particles of PVX-coded movement protein, the product of the first gene of triple gene block (TGBp1). Here we report that a similar effect occurs via in situ phosphorylation of the PVX coat protein (CP) by Ser/Thr protein kinase (PK) C, the mixture of casein kinases I and II or by cytoplasmic PK(s) from Nicotiana glutinosa leaves. Immunochemical analyses indicated that phosphorylation induced conformational changes in PVX CP. The N-terminal region of the PVX CP, rich in Ser and Thr residues, is exposed at the virion surface and can be removed by treatment with trypsin. We showed that (i) trypsin treatment removed the bulk of (32)P-radioactivity from in situ phosphorylated PVX CP, (ii) PVX containing N-terminally truncated CP (PVX-Ptd) failed to be translationally activated by phosphorylation, and (iii) the specific infectivity of PVX-Ptd was reduced. However, the PVX-Ptd RNA remained intact and PVX-Ptd could be translationally activated by the PVX MP TGBp1. We hypothesize that phosphorylation of the parental PVX by cytoplasmic PK(s) in vivo renders PVX RNA translatable in primary inoculated cells, whereas translational activation of the progeny virions destined for plasmodesmata trafficking is triggered by TGBp1.

show abstract

Anomalous electrophoretic behavior of the major potato virus X RNA translation product

Cited by 6 publications

References 12 publications

Examination of potato virus X proteins synthesized in infected tobacco plants

Examination of potato virus X proteins synthesized in infected tobacco plants

The Movement Protein-Triggered in Situ Conversion of Potato Virus X Virion RNA from a Nontranslatable into a Translatable Form

Translational Activation of Encapsidated Potato Virus X RNA by Coat Protein Phosphorylation

Contact Info

Product

Resources

About