Hyeong Cheol Park*" /> Wonkyun Choi" /> Hyeong Cheol Park*, Wonkyun Choi, Hee Jin Park,Mi Sun Cheong, Yoon Duck Koo, Gilok Shin, Woo Sik Chung, Woe-Yeon Kim, Min Gab Kim, Ray A. Bressan, Hans J. Bohnert, Sang Yeol Lee, and Dae-Jin Yun*" /> Hyeong Cheol Park*, Wonkyun Choi, Hee Jin Park,Mi Sun Cheong, Yoon Duck Koo, Gilok Shin, Woo Sik Chung, Woe-Yeon Kim, Min Gab Kim, Ray A. Bressan, Hans J. Bohnert, Sang Yeol Lee, and Dae-Jin Yun*. Mol. Cells 2011;32:143-51. https://doi.org/10.1007/s10059-011-2297-3">
Top

Research Article

Split Viewer

Mol. Cells 2011; 32(2): 143-151

Published online August 31, 2011

https://doi.org/10.1007/s10059-011-2297-3

© The Korean Society for Molecular and Cellular Biology

Identification and Molecular Properties of SUMO-Binding Proteins in Arabidopsis

Hyeong Cheol Park1,6,*, Wonkyun Choi1,6, Hee Jin Park1,2,6, Mi Sun Cheong1, Yoon Duck Koo1, Gilok Shin1, Woo Sik Chung1, Woe-Yeon Kim1, Min Gab Kim3, Ray A. Bressan1,2,4, Hans J. Bohnert1,5, Sang Yeol Lee1, and Dae-Jin Yun1,*

1Division of Applied Life Science (Brain Korea 21 Program), and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju 660-701, Korea, 2Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana, USA, 3Bio-Crops Development Division, Department of Agricultural Biotechnology, National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Korea, 4King Abdullah University of Science and Technology, Thuwal, 23955-6900, Saudi Arabia, 5Departments of Plant Biology and of Crop Sciences, University of Illinois at Urbana-Champaign, Urbana, USA, 6These authors contributed equally to this work.

Correspondence to : *Correspondence: hcpark@gnu.ac.kr (HCP); djyun@gnu.ac.kr (DJY)

Received: December 2, 2011; Revised: April 20, 2011; Accepted: April 28, 2011

Abstract

Reversible conjugation of the small ubiquitin modifier (SUMO) peptide to proteins (SUMOylation) plays important roles in cellular processes in animals and yeasts. However, little is known about plant SUMO targets. To identify SUMO substrates in Arabidopsis and to probe for biological functions of SUMO proteins, we constructed 6xHis-3xFLAG fused AtSUMO1 (HFAtSUMO1) controlled by the CaMV35S promoter for transformation into Arabidopsis Col-0. After heat treatment, an increased sumoylation pattern was de-tected in the transgenic plants. SUMO1-modified proteins were selected after two-dimensional gel electrophoresis (2-DE) image analysis and identified using matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). We identified 27 proteins involved in a variety of processes such as nucleic acid metabolism, signaling, metabolism, and including proteins of unknown functions. Binding and sumoylation patterns were confirmed independently. Surprisingly, MCM3 (At5G46280), a DNA replication licensing factor, only interacted with and became sumoylated by AtSUMO1, but not by SUMO1?GG or AtSUMO3. The results suggest specific interactions between sumoylation targets and particular sumoylation enzymes.

Keywords Arabidopsis, mass spectrometry, proteomics, SUMO binding proteins, Sumoylation

Article

Research Article

Mol. Cells 2011; 32(2): 143-151

Published online August 31, 2011 https://doi.org/10.1007/s10059-011-2297-3

Copyright © The Korean Society for Molecular and Cellular Biology.

Identification and Molecular Properties of SUMO-Binding Proteins in Arabidopsis

Hyeong Cheol Park1,6,*, Wonkyun Choi1,6, Hee Jin Park1,2,6, Mi Sun Cheong1, Yoon Duck Koo1, Gilok Shin1, Woo Sik Chung1, Woe-Yeon Kim1, Min Gab Kim3, Ray A. Bressan1,2,4, Hans J. Bohnert1,5, Sang Yeol Lee1, and Dae-Jin Yun1,*

1Division of Applied Life Science (Brain Korea 21 Program), and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju 660-701, Korea, 2Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana, USA, 3Bio-Crops Development Division, Department of Agricultural Biotechnology, National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Korea, 4King Abdullah University of Science and Technology, Thuwal, 23955-6900, Saudi Arabia, 5Departments of Plant Biology and of Crop Sciences, University of Illinois at Urbana-Champaign, Urbana, USA, 6These authors contributed equally to this work.

Correspondence to:*Correspondence: hcpark@gnu.ac.kr (HCP); djyun@gnu.ac.kr (DJY)

Received: December 2, 2011; Revised: April 20, 2011; Accepted: April 28, 2011

Abstract

Reversible conjugation of the small ubiquitin modifier (SUMO) peptide to proteins (SUMOylation) plays important roles in cellular processes in animals and yeasts. However, little is known about plant SUMO targets. To identify SUMO substrates in Arabidopsis and to probe for biological functions of SUMO proteins, we constructed 6xHis-3xFLAG fused AtSUMO1 (HFAtSUMO1) controlled by the CaMV35S promoter for transformation into Arabidopsis Col-0. After heat treatment, an increased sumoylation pattern was de-tected in the transgenic plants. SUMO1-modified proteins were selected after two-dimensional gel electrophoresis (2-DE) image analysis and identified using matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). We identified 27 proteins involved in a variety of processes such as nucleic acid metabolism, signaling, metabolism, and including proteins of unknown functions. Binding and sumoylation patterns were confirmed independently. Surprisingly, MCM3 (At5G46280), a DNA replication licensing factor, only interacted with and became sumoylated by AtSUMO1, but not by SUMO1?GG or AtSUMO3. The results suggest specific interactions between sumoylation targets and particular sumoylation enzymes.

Keywords: Arabidopsis, mass spectrometry, proteomics, SUMO binding proteins, Sumoylation

Mol. Cells
Jun 30, 2022 Vol.45 No.6, pp. 353~434
COVER PICTURE
ERα is modified by UFM1 and this modification (ufmylation) plays a crucial role in promoting the stability of ERα and breast cancer development. However, when ERα is deufmylated and then ubiquitinated, it disappears by proteasome-mediated degradation (Yoo et al., pp. 425-434).

Share this article on

  • line
  • mail

Related articles in Mol. Cells

Molecules and Cells

eISSN 0219-1032
qr-code Download