The location of the 5' end of the potato leafroll luteovirus subgenomic coat protein mRNA

Miller, Judy; Mayo, M. A.

doi:10.1099/0022-1317-72-11-2633

Cited by 44 publications

(25 citation statements)

References 28 publications

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“…Two oligonucleotides denoted primer 3 (complementary to nt 407 to 426) and primer 4 (complementary to nt 1521 to 1540) of RNA3, respectively, were utilized to determine the 5' ends of both RNA4 and the subgenomic RNA coding for the coat protein. Extension of 5'-32P-labelled primers was carried out as described by Miller & Mayo (1991), using TNA from infected N. benthamiana and purified virus RNA as template. The products of primer extension were analysed by electrophoresis in an 8 % polyacrylamide gel containing 7 M-urea.…”

Section: Methodsmentioning

confidence: 99%

The nucleotide sequence of RNA3 and RNA4 of olive latent virus 2

Grieco¹,

Martelli²,

Savino³

1995

Journal of General Virology

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Olive latent virus 2 (OLV2), a virus with particle shapes resembling those of alfalfa mosaic alfamovirus (AMV), has four major R N A species, two of which (RNA3 and RNA4) were completely sequenced. RNA3 was a bicistronic molecule containing two clear-cut ORFs, one of which (ORF1) coded for a 36-5 kDa polypeptide with conserved motifs of the ' 3 0 K superfamily' movement proteins and the other (ORF2) encoded a 20 kDa polypeptide identified as the viral coat protein. RNA4, which was a little smaller than RNA3 (2078 nt versus 2438 nt), also differed from RNA3 in a few positions, but its in vitro translation did not produce any protein. By contrast, an additional RNA, 1042 nt in size with strong sequence homology with RNA3 and RNA4, was identified in RNA extracts from infected plants. This RNA, which may be a subgenomic form of RNA3 carrying the coat protein cistron, is apparently encapsidated to a very small extent, or not at all. Comparative computer-assisted analysis of virus-coded protein sequences disclosed homologies suggesting that OLV2 may belong to the family Bromoviridae, but as an entity separated from the currently known genera.

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Section: Methodsmentioning

confidence: 99%

The nucleotide sequence of RNA3 and RNA4 of olive latent virus 2

Grieco¹,

Martelli²,

Savino³

1995

Journal of General Virology

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“…In addition to these six ORFs, two small ORFs were identified within the 39-terminal region (Ashoub et al, 1998). Jaag et al (2003) reported that another small ORF within the P1 region, in an alternative reading frame, was essential for viral multiplication.Like many positive-strand ssRNA viruses, PLRV has been found to express the products of its genome in a variety of ways, including transcription of subgenomic mRNAs, translational frameshifting, stop-codon readthrough, internal initiation, translation initiation at an internal ribosomal entry site and polyprotein processing (Miller et al, 1995;Mayo & Ziegler-Graff, 1996).Proteins encoded within the 59-proximal region are translated from the genomic RNA, whereas 39-proximal ORFs are expressed from two subgenomic RNAs (Tacke et al, 1990;Miller & Mayo, 1991;Ashoub et al, 1998). ORF2, which encodes the RNA-dependent RNA polymerase, is expressed via 21 frameshifting at a site within ORF1 (Prüfer et al, 1992).…”

mentioning

confidence: 99%

“…Proteins encoded within the 59-proximal region are translated from the genomic RNA, whereas 39-proximal ORFs are expressed from two subgenomic RNAs (Tacke et al, 1990;Miller & Mayo, 1991;Ashoub et al, 1998). ORF2, which encodes the RNA-dependent RNA polymerase, is expressed via 21 frameshifting at a site within ORF1 (Prüfer et al, 1992).…”

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confidence: 99%

A novel cleavage site within the potato leafroll virus P1 polyprotein

Halpin

Ryan

2007

Journal of General Virology

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To study the proteolytic processing of the potato leafroll virus replicase proteins, the multidomain P1 protein with a c-myc epitope tag attached at the N terminus was expressed in insect cells by using the baculovirus system. Western blotting showed that P1 was cleaved at a site upstream of the serine protease domain, in addition to the cleavage site downstream of the protease domain. Mutational analysis showed that the serine protease domain within P1 was responsible for this cleavage. To characterize this novel cleavage site further, a portion of the P1 protein comprising the protease domain and the two cleavage sites was expressed in Escherichia coli. A similar cleavage event was observed in bacteria and was abolished when the P1 protease was inactivated by mutation. Peptide-sequencing studies indicated that this cleavage occurred at a Glu/Arg junction, separating the N-terminal 204 residues from the serine protease domain of P1.Potato leafroll virus (PLRV) is now classified as the type member of the genus Polerovirus of the family Luteoviridae, which also contains the genera Luteovirus and Enamovirus (Pringle, 1998). PLRV virions appear to be non-enveloped, isometric particles with a diameter of 24 nm (Harrison, 1984). Viral particles contain a single-stranded (ss), positive-sense genomic RNA of some 5.8 kb in length (Mayo et al., 1989;van der Wilk et al., 1989;Keese et al., 1990). Like most luteoviruses, PLRV genomic RNA encodes six major open reading frames (ORFs), which are arranged into two blocks separated by a small intergenic region. ORF0 encodes a 28 kDa protein that is important for virus accumulation (Sadowy et al., 2001b). Site-directed mutagenetic analysis has shown that P1 and P2, encoded by ORF1 and ORF2, are required for virus replication (Sadowy et al., 2001a) -consistent with studies on beet western yellows virus (BWYV), another polerovirus (Reutenauer et al., 1993). The 39-terminal block encodes the capsid protein (ORF3), the putative movement protein (ORF4) and the aphid transmission protein (ORF5; reviewed by Miller et al., 1995;Mayo & Ziegler-Graff, 1996). In addition to these six ORFs, two small ORFs were identified within the 39-terminal region (Ashoub et al., 1998). Jaag et al. (2003) reported that another small ORF within the P1 region, in an alternative reading frame, was essential for viral multiplication.Like many positive-strand ssRNA viruses, PLRV has been found to express the products of its genome in a variety of ways, including transcription of subgenomic mRNAs, translational frameshifting, stop-codon readthrough, internal initiation, translation initiation at an internal ribosomal entry site and polyprotein processing (Miller et al., 1995;Mayo & Ziegler-Graff, 1996).Proteins encoded within the 59-proximal region are translated from the genomic RNA, whereas 39-proximal ORFs are expressed from two subgenomic RNAs (Tacke et al., 1990;Miller & Mayo, 1991;Ashoub et al., 1998). ORF2, which encodes the RNA-dependent RNA polymerase, is expressed via 21 frameshifting at a site withi...

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“…The 17 kDa protein (prl7) of PLRV is encoded by ORF4 in the 3' half of the viral genome. It is translated from subgenomic PLRV RNA1 (sgRNAl) by internal translation initiation at an in vitro translation efficiency which is sevenfold higher as compared to that of the coat protein, the main structural protein of the PLRV virion, which is translated from the identical sgRNAl [4,5].…”

Section: Introductionmentioning

confidence: 99%

The potato leafroll virus 17K movement protein is phosphorylated by a membrane‐associated protein kinase from potato with biochemical features of protein kinase C

Sokolova¹,

Prüfer²,

Tacke³

et al. 1997

FEBS Letters

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In planta the protein is predominantly present in its phosphorylated form, but it is rapidly dephosphorylated during isolation under native conditions. In an effort to examine the nature of the protein kinase(s) involved in the phosphorylation reaction, prl7 deletion mutants were expressed as fusion proteins in a bacterial expression vector system and tested for their ability to be phosphorylated by potato membrane preparations as well as by commercially available kinases. A fusion protein containing the nucleic acid-binding, basic, C-proximal domain (prl7Cl) was identified to be phosphorylated by a Ca 2+ -and phospholipiddependent, membrane-associated protein kinase. This protein kinase activity was inhibited by the addition of (19-36) protein kinase C (PKC) inhibitory peptide, known to be a highly specific inhibitor of mammalian PKC. Moreover, also the mammalian PKC from rat was able to phosphorylate prl7 in vitro. The results suggest that phosphorylation of prl7 takes place at membranous structures, possibly at the deltoid plasmodesmata connecting the sieve cell-companion cell complex of the phloem, by the activity of PKC-related, membrane-associated protein kinase activity.

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The location of the 5' end of the potato leafroll luteovirus subgenomic coat protein mRNA

Cited by 44 publications

References 28 publications

The nucleotide sequence of RNA3 and RNA4 of olive latent virus 2

The nucleotide sequence of RNA3 and RNA4 of olive latent virus 2

A novel cleavage site within the potato leafroll virus P1 polyprotein

The potato leafroll virus 17K movement protein is phosphorylated by a membrane‐associated protein kinase from potato with biochemical features of protein kinase C

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