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Analysis of potyvirus terminal protein VPg-transgenic Arabidopsis thaliana plants

100%

Wojtal I., Piontek P., Grzela R., Jarmołowski A., Zagórski W., Chroboczek J.

2011

vol. 58

issue 3

349-353

Virus-coded VPg protein of Potato virus Y (PVY) does not have homologs apart from other VPgs. Since VPg is indispensable for the potyvirus life cycle, it appeared a good candidate for eliciting pathogen-derived resistance to PVY. Following agroinfection used to obtain PVY VPg-transgenic Arabidopsis thaliana plants, only few transgenic seeds were recovered giving rise to six transgenic plants that contained the VPg gene with the correct sequence. They generated VPg mRNA, but VPg protein was not detected. Some plants were immune to PVY infection suggesting post-transcriptional gene silencing. However, the likely PVY VPg toxicity exerted at an early stage of transformed seeds development precludes its use for engineering pathogen-derived resistance.

Certain protein transducing agents convert translocated proteins into cell killers

84%

Tcherniuk S., Fiser A.-L., Derouazi M., Toussaint B., Wang Y., Wojtal I., Kondo E., Szolajska E., Chroboczek J.

2012

vol. 59

issue 3

433-439

The majority of proteins are unable to translocate into the cell interior. Hence for peptide- and protein-based therapeutics a direct intracytoplasmic delivery with the aid of transducing agents is an attractive approach. We wanted to deliver to the cell interior a putatively cytotoxic protein VPg. Protein transduction was achieved in vitro with three different commercial products. However, in our hands, delivery of various control proteins without known deleterious effects, as well as of protein VPg, always induced cell death. Finally, we used a novel transducing peptide Wr-T, which was not toxic to cultured cells, even in a quite large range of concentrations. Most importantly, control protein delivered to cells in culture did not display any toxicity while VPg protein exerted a strong cytotoxic effect. These data show that results obtained with cell-penetrating agents should be interpreted with caution.

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2 Acta Biochimica Polonica

1 2012

1 2011

2 Chroboczek J.

2 Wojtal I.

1 Derouazi M.

1 Fiser A.-L.

1 Grzela R.

1 Jarmołowski A.

1 Kondo E.

1 Piontek P.

1 Szolajska E.

1 Tcherniuk S.

1 Toussaint B.

1 Wang Y.

1 Zagórski W.

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Analysis of potyvirus terminal protein VPg-transgenic Arabidopsis thaliana plants

Certain protein transducing agents convert translocated proteins into cell killers