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PMID:22511869

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Citation

Vijayapalani, P, Maeshima, M, Nagasaki-Takekuchi, N and Miller, WA (2012) Interaction of the trans-frame potyvirus protein P3N-PIPO with host protein PCaP1 facilitates potyvirus movement. PLoS Pathog. 8:e1002639

Abstract

A small open reading frame (ORF), pipo, overlaps with the P3 coding region of the potyviral polyprotein ORF. Previous evidence suggested a requirement for pipo for efficient viral cell-to-cell movement. Here, we provide immunoblotting evidence that the protein PIPO is expressed as a trans-frame protein consisting of the amino-terminal half of P3 fused to PIPO (P3N-PIPO). P3N-PIPO of Turnip mosaic virus (TuMV) fused to GFP facilitates its own cell-to-cell movement. Using a yeast two-hybrid screen, co-immunoprecipitation assays, and bimolecular fluorescence complementation (BiFC) assays, we found that P3N-PIPO interacts with host protein PCaP1, a cation-binding protein that attaches to the plasma membrane via myristoylation. BiFC revealed that it is the PIPO domain of P3N-PIPO that binds PCaP1 and that myristoylation of PCaP1 is unnecessary for interaction with P3N-PIPO. In PCaP1 knockout mutants (pcap1) of Arabidopsis, accumulation of TuMV harboring a GFP gene (TuMV-GFP) was drastically reduced relative to the virus level in wild-type plants, only small localized spots of GFP were visible, and the plants showed few symptoms. In contrast, TuMV-GFP infection in wild-type Arabidopsis yielded large green fluorescent patches, and caused severe stunting. However, viral RNA accumulated to high level in protoplasts from pcap1 plants indicating that PCaP1 is not required for TuMV RNA synthesis. In contrast to TuMV, the tobamovirus Oilseed rape mosaic virus did not require PCaP1 to infect Arabidopsis plants. We conclude that potyviral P3N-PIPO interacts specifically with the host plasma membrane protein PCaP1 to participate in cell-to-cell movement. We speculate that PCaP1 links a complex of viral proteins and genomic RNA to the plasma membrane by binding P3N-PIPO, enabling localization to the plasmodesmata and cell-to-cell movement. The PCaP1 knockout may contribute to a new strategy for recessive resistance to potyviruses.

Links

PubMed PMC3325209 Online version:10.1371/journal.ppat.1002639

Keywords

Arabidopsis/genetics; Arabidopsis/metabolism; Arabidopsis/virology; Arabidopsis Proteins/genetics; Arabidopsis Proteins/metabolism; Carrier Proteins/genetics; Carrier Proteins/metabolism; Cell Membrane/genetics; Cell Membrane/metabolism; Cell Membrane/virology; Gene Knockdown Techniques; Genome, Viral/physiology; Open Reading Frames/physiology; Plant Diseases/virology; Plants, Genetically Modified; Potyvirus/genetics; Potyvirus/metabolism; RNA, Viral/genetics; RNA, Viral/metabolism; Saccharomyces cerevisiae; Two-Hybrid System Techniques; Viral Proteins/genetics; Viral Proteins/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

TUMVJ:MVP

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q96262

F

Seeded From UniProt

complete

ARATH:PCAP1

located_in

GO:0009506: plasmodesma

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

ARATH:PCAP1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P0CK11

F

Seeded From UniProt

complete

ARATH:PCAP1

acts_upstream_of_or_within

GO:0075733: intracellular transport of virus

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:Q96262

involved_in

GO:0075733: intracellular transport of virus

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:Q96262

part_of

GO:0009506: plasmodesma

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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