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Mol. Cells 2012; 33(4): 379-384

Published online April 30, 2012

https://doi.org/10.1007/s10059-012-2298-x

© The Korean Society for Molecular and Cellular Biology

Identification of Tobacco Proteins Associated with the Stem-Loop 1 RNAs of Potato virus X

Sang-Yun Cho1,3, Won Kyong Cho1,3, and Kook-Hyung Kim1,2,*

1Department of Agricultural Biotechnology and Plant Genomics and Breeding Institute, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea, 2Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea, 3These authors contributed equally to this work.

Correspondence to : *Correspondence: kookkim@snu.ac.kr

Received: December 30, 2011; Revised: January 28, 2012; Accepted: February 1, 2012

Abstract

Potato virus X (PVX) contains five viral proteins as well as cis-acting elements like stem-loop 1 (SL1) RNAs at the 5? region. SL1 RNAs are involved in PVX RNA replication, encapsidation, translation, and cell-to-cell movement. In this study, we performed two-dimensional electrophoresis Northwestern blot analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry and identified 24 tobacco proteins that interact with SL1 RNAs. Interestingly, one-third of the identified host proteins have been shown to interact with many plant viral proteins. In addition, we demonstrated that PVX capsid protein can bind to both SL1(+/-) RNAs. We further selected three Nicotiana benthamiana proteins including NbMPB2Cb, NbMBF1, and NbCPIP2a, to confirm results of Northwestern blot analysis. Electrophoretic mobility shift assay showed that NbMPB2Cb and NbMBF1 bind to both SL1(+/-) RNAs in vitro. In contrast, NbCPIP2a interacts only SL1(+) RNA. Taken together, we provide a list of host proteins interacting with PVX SL1 RNAs, which would be good candidates for the study of viral RNA-host protein interaction.

Keywords host protein, northwestern blot, PVX, RNA-protein interaction, SL1 RNAs

Article

Research Article

Mol. Cells 2012; 33(4): 379-384

Published online April 30, 2012 https://doi.org/10.1007/s10059-012-2298-x

Copyright © The Korean Society for Molecular and Cellular Biology.

Identification of Tobacco Proteins Associated with the Stem-Loop 1 RNAs of Potato virus X

Sang-Yun Cho1,3, Won Kyong Cho1,3, and Kook-Hyung Kim1,2,*

1Department of Agricultural Biotechnology and Plant Genomics and Breeding Institute, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea, 2Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea, 3These authors contributed equally to this work.

Correspondence to:*Correspondence: kookkim@snu.ac.kr

Received: December 30, 2011; Revised: January 28, 2012; Accepted: February 1, 2012

Abstract

Potato virus X (PVX) contains five viral proteins as well as cis-acting elements like stem-loop 1 (SL1) RNAs at the 5? region. SL1 RNAs are involved in PVX RNA replication, encapsidation, translation, and cell-to-cell movement. In this study, we performed two-dimensional electrophoresis Northwestern blot analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry and identified 24 tobacco proteins that interact with SL1 RNAs. Interestingly, one-third of the identified host proteins have been shown to interact with many plant viral proteins. In addition, we demonstrated that PVX capsid protein can bind to both SL1(+/-) RNAs. We further selected three Nicotiana benthamiana proteins including NbMPB2Cb, NbMBF1, and NbCPIP2a, to confirm results of Northwestern blot analysis. Electrophoretic mobility shift assay showed that NbMPB2Cb and NbMBF1 bind to both SL1(+/-) RNAs in vitro. In contrast, NbCPIP2a interacts only SL1(+) RNA. Taken together, we provide a list of host proteins interacting with PVX SL1 RNAs, which would be good candidates for the study of viral RNA-host protein interaction.

Keywords: host protein, northwestern blot, PVX, RNA-protein interaction, SL1 RNAs

Mol. Cells
Sep 30, 2022 Vol.45 No.9, pp. 603~672
COVER PICTURE
The Target of Rapamycin Complex (TORC) is a central regulatory hub in eukaryotes, which is well conserved in diverse plant species, including tomato (Solanum lycopersicum). Inhibition of TORC genes (SlTOR, SlLST8, and SlRAPTOR) by VIGS (virus-induced gene silencing) results in early fruit ripening in tomato. The red/ orange tomatoes are early-ripened TORC-silenced fruits, while the green tomato is a control fruit. Top, left, control fruit (TRV2-myc); top, right, TRV2-SlLST8; bottom, left, TRV2-SlTOR; bottom, right, TRV2-SlRAPTOR(Choi et al., pp. 660-672).

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