The host Pex19p plays a role in peroxisomal localization of tombusvirus replication proteins

Kunj B. Pathak; Zsuzsanna Sasvari; Peter D. Nagy

doi:10.1016/j.virol.2008.06.044

The host Pex19p plays a role in peroxisomal localization of tombusvirus replication proteins

Kunj B. Pathak, Zsuzsanna Sasvari, Peter D. Nagy

Plant Pathology

Research output: Contribution to journal › Article › peer-review

88 Scopus citations

Abstract

Replication of Tomato bushy stunt virus (TBSV) RNA takes place on the cytosolic membrane surface of peroxisomes in plants and in yeast, a model host. To identify the host proteins involved in assisting the peroxisomal localization of the tombusvirus p33 replication protein, we tested if p33 could bind directly to yeast proteins involved in peroxisomal transport in vitro. This work has led to the demonstration of Pex19p-p33 interaction via pull-down and co-purification experiments. Pex19p was also detected in the tombusvirus replicase after protein cross-linking, suggesting that Pex19p transiently binds to the replicase as could be expected from a transporter. To validate the importance of Pex19p-p33 interaction in TBSV replication in yeast, we re-targeted Pex19p to the mitochondria, which resulted in the re-distribution of a large fraction of p33 to the mitochondria. The expression of the mitochondrial-targeted Pex19p inhibited TBSV RNA accumulation by 2-4-fold in vivo and reduced the in vitro activity of the tombusvirus replicase by 80%. These data support the model that Pex19p is a cellular transporter for localization of p33 replication protein to the host peroxisomal membranes.

Original language	English
Pages (from-to)	294-305
Number of pages	12
Journal	Virology
Volume	379
Issue number	2
DOIs	https://doi.org/10.1016/j.virol.2008.06.044
State	Published - Sep 30 2008

Bibliographical note

Funding Information:
The authors thank Dr. Judit Pogany and Kai Xu for valuable comments. Dr. Robert Wang for his help with Confocal laser microscopy and Dr. Li for his help with the co-purification experiments. The authors are grateful to Drs. R. Sopko and B. Andrews, University of Toronto, Ontario, Canada for the yeast GST-ORF over-expression library, to Dr. R. Erdmann, Bochum, Germany for the anti-Pex19 antibody. This work was supported by the National Science Foundation (IOB-0517218), and the Kentucky Tobacco Research and Development Center. KBP was also supported by Philip Morris fellowship.

Keywords

Dominant negative
Host-virus interaction
Peroxisome
RNA virus
Replication
Tomato bushy stunt virus
Tombusvirus
Yeast

ASJC Scopus subject areas

Virology

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10.1016/j.virol.2008.06.044

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title = "The host Pex19p plays a role in peroxisomal localization of tombusvirus replication proteins",

abstract = "Replication of Tomato bushy stunt virus (TBSV) RNA takes place on the cytosolic membrane surface of peroxisomes in plants and in yeast, a model host. To identify the host proteins involved in assisting the peroxisomal localization of the tombusvirus p33 replication protein, we tested if p33 could bind directly to yeast proteins involved in peroxisomal transport in vitro. This work has led to the demonstration of Pex19p-p33 interaction via pull-down and co-purification experiments. Pex19p was also detected in the tombusvirus replicase after protein cross-linking, suggesting that Pex19p transiently binds to the replicase as could be expected from a transporter. To validate the importance of Pex19p-p33 interaction in TBSV replication in yeast, we re-targeted Pex19p to the mitochondria, which resulted in the re-distribution of a large fraction of p33 to the mitochondria. The expression of the mitochondrial-targeted Pex19p inhibited TBSV RNA accumulation by 2-4-fold in vivo and reduced the in vitro activity of the tombusvirus replicase by 80%. These data support the model that Pex19p is a cellular transporter for localization of p33 replication protein to the host peroxisomal membranes.",

keywords = "Dominant negative, Host-virus interaction, Peroxisome, RNA virus, Replication, Tomato bushy stunt virus, Tombusvirus, Yeast",

author = "Pathak, {Kunj B.} and Zsuzsanna Sasvari and Nagy, {Peter D.}",

note = "Funding Information: The authors thank Dr. Judit Pogany and Kai Xu for valuable comments. Dr. Robert Wang for his help with Confocal laser microscopy and Dr. Li for his help with the co-purification experiments. The authors are grateful to Drs. R. Sopko and B. Andrews, University of Toronto, Ontario, Canada for the yeast GST-ORF over-expression library, to Dr. R. Erdmann, Bochum, Germany for the anti-Pex19 antibody. This work was supported by the National Science Foundation (IOB-0517218), and the Kentucky Tobacco Research and Development Center. KBP was also supported by Philip Morris fellowship.",

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doi = "10.1016/j.virol.2008.06.044",

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AU - Pathak, Kunj B.

AU - Sasvari, Zsuzsanna

AU - Nagy, Peter D.

N1 - Funding Information: The authors thank Dr. Judit Pogany and Kai Xu for valuable comments. Dr. Robert Wang for his help with Confocal laser microscopy and Dr. Li for his help with the co-purification experiments. The authors are grateful to Drs. R. Sopko and B. Andrews, University of Toronto, Ontario, Canada for the yeast GST-ORF over-expression library, to Dr. R. Erdmann, Bochum, Germany for the anti-Pex19 antibody. This work was supported by the National Science Foundation (IOB-0517218), and the Kentucky Tobacco Research and Development Center. KBP was also supported by Philip Morris fellowship.

PY - 2008/9/30

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N2 - Replication of Tomato bushy stunt virus (TBSV) RNA takes place on the cytosolic membrane surface of peroxisomes in plants and in yeast, a model host. To identify the host proteins involved in assisting the peroxisomal localization of the tombusvirus p33 replication protein, we tested if p33 could bind directly to yeast proteins involved in peroxisomal transport in vitro. This work has led to the demonstration of Pex19p-p33 interaction via pull-down and co-purification experiments. Pex19p was also detected in the tombusvirus replicase after protein cross-linking, suggesting that Pex19p transiently binds to the replicase as could be expected from a transporter. To validate the importance of Pex19p-p33 interaction in TBSV replication in yeast, we re-targeted Pex19p to the mitochondria, which resulted in the re-distribution of a large fraction of p33 to the mitochondria. The expression of the mitochondrial-targeted Pex19p inhibited TBSV RNA accumulation by 2-4-fold in vivo and reduced the in vitro activity of the tombusvirus replicase by 80%. These data support the model that Pex19p is a cellular transporter for localization of p33 replication protein to the host peroxisomal membranes.

AB - Replication of Tomato bushy stunt virus (TBSV) RNA takes place on the cytosolic membrane surface of peroxisomes in plants and in yeast, a model host. To identify the host proteins involved in assisting the peroxisomal localization of the tombusvirus p33 replication protein, we tested if p33 could bind directly to yeast proteins involved in peroxisomal transport in vitro. This work has led to the demonstration of Pex19p-p33 interaction via pull-down and co-purification experiments. Pex19p was also detected in the tombusvirus replicase after protein cross-linking, suggesting that Pex19p transiently binds to the replicase as could be expected from a transporter. To validate the importance of Pex19p-p33 interaction in TBSV replication in yeast, we re-targeted Pex19p to the mitochondria, which resulted in the re-distribution of a large fraction of p33 to the mitochondria. The expression of the mitochondrial-targeted Pex19p inhibited TBSV RNA accumulation by 2-4-fold in vivo and reduced the in vitro activity of the tombusvirus replicase by 80%. These data support the model that Pex19p is a cellular transporter for localization of p33 replication protein to the host peroxisomal membranes.

KW - Dominant negative

KW - Host-virus interaction

KW - Peroxisome

KW - RNA virus

KW - Replication

KW - Tomato bushy stunt virus

KW - Tombusvirus

KW - Yeast

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M3 - Article

C2 - 18684480

AN - SCOPUS:50349103141

SN - 0042-6822

VL - 379

SP - 294

EP - 305

JO - Virology

JF - Virology

IS - 2

ER -

The host Pex19p plays a role in peroxisomal localization of tombusvirus replication proteins

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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