TGBp3 triggers the unfolded protein response and SKP1-dependent programmed cell death
Version of Record online: 12 NOV 2012
© 2012 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2012 BSPP AND BLACKWELL PUBLISHING LTD
Molecular Plant Pathology
Volume 14, Issue 3, pages 241–255, April 2013
How to Cite
YE, C.-M., Chen, S., Payton, M., Dickman, M. B. and Verchot, J. (2013), TGBp3 triggers the unfolded protein response and SKP1-dependent programmed cell death. Molecular Plant Pathology, 14: 241–255. doi: 10.1111/mpp.12000
- Issue online: 4 MAR 2013
- Version of Record online: 12 NOV 2012
- US Department of Agriculture National Research Initiative (USDA NRI). Grant Number: OKL02649
Fig. S1 (A) qRT-PCR analyzing bZIP60, BiP and SKP1 transcript abundance. The plots show the relative abundance of transcripts due to each TMV, PVX, and TMV plus PVX infection. Clearly transcript abundance is higher during PVX infection. Furthermore, TMV has a dominant suppressive effect on the ability of PVX to induce host gene expression. Furthermore, we fused the PVX replicase, TGBp1, and CP to the CaMV35S promoter and delivered these plasmids to plants using agro-infiltration. qRT-PCR shows host gene expression is not induced by these factors. Immunoblots demonstrating the relative abundance of TGBp1 (B), PVX CP (D), and commassie stained gel detecting TGBp2 (C) when expressed from either PVX or TMV vector in infected N. benthamiana leaves. Since we lack antisera to TGBp2, commassie staining was used to identify a band that likely corresponds to TGBp2 in infected leaf extracts. Commassie stained immunoblot membranes in panels B and D show similar levels of protein in each lane.
Fig. S2 Immunoblot analysis detecting PVX TGBp3-His protein using antisera detecting the His tag. (A) Leaf extracts from mock inoculated (M; lane 1), PVX-TGBp3His (lanes 2–5), TMV-p3His (lanes 6–9), and agrobacterium (lane 11) expressing TGBp3His from CaMV 35S promoter (lanes 12–13). Leaf extracts were taken from virus-inoculated (Inoc) and systemically (Syst) infected tissues. TMV-p3His showed higher levels of protein accumulation than PVX-TGBp3His and required 20 s exposure to film to see abundance of protein in the top panel. To visualize TGBp3His expressed from PVX, we exposed the film for 1 min and this is shown in the middle panel. TGBp3His expression from Agrobacterium was comparable to TMV-p3His as seen after 20 s exposure to film. Below is a commassie stained PVDF membrane photographed after electrophoretic transfer to show protein levels in each lane were comparable. (B) Immunoblot detects TMV coat protein. TMV-p3 and TMV-p3His were detected in systemic tissues and produced comparable levels of TMV coat protein indicating that the His tag is not deleterious to virus infection.
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