Identification and molecular properties of SUMO-binding proteins in arabidopsis

Hyeongcheol Park; Wonkyun Choi; Heejin Park; Misun Cheong; Yoonduck Koo; Gilok Shin; Woosik Chung; Woeyeon Kim; Mingab Kim; Ray Anthony Bressan; Hans Jürgen Bohnert; Sangyeol Lee; Daejin Yun

doi:10.1007/s10059-011-2297-3

Identification and molecular properties of SUMO-binding proteins in arabidopsis

Hyeongcheol Park, Wonkyun Choi, Heejin Park, Misun Cheong, Yoonduck Koo, Gilok Shin, Woosik Chung, Woeyeon Kim, Mingab Kim, Ray Anthony Bressan, Hans Jürgen Bohnert, Sangyeol Lee, Daejin Yun

Biological, Environmental Sciences and Engineering

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

40 Scopus citations

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 detected 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 SUMO1ΔGG or AtSUMO3. The results suggest specific interactions between sumoylation targets and particular sumoylation enzymes. ©2011 KSMCB.

Original language	English (US)
Pages (from-to)	143-151
Number of pages	9
Journal	Molecules and Cells
Volume	32
Issue number	2
DOIs	https://doi.org/10.1007/s10059-011-2297-3
State	Published - May 20 2011

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: We thank Dr. Jian-Min Zhou for providing the NLuc and CLuc plasmids and Dr. Katsunori Tanaka for providing the pCDFDuet-AtSUMO-AtSCE1, pACYCDuet-AtSAEb-AtSAE2, and pET28a-AtMYB30 plasmids. This work was supported by grants from the Biogreen 21 Program (grant No. PJ006654) of the Rural Development Administration, World Class University Program (R32-10148) funded by the Ministry of Education, Science and Technology in Korea, and the National Research Foundation of Korea Grant funded by the Korean Government (Ministry of Education, Science and Technology) [NRF-2010-359-F00006]. GS was supported by scholarship from the Brain Korea 21 program of the Ministry of Education, Science and Technology in Korea.

ASJC Scopus subject areas

Cell Biology
Molecular Biology

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10.1007/s10059-011-2297-3

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title = "Identification and molecular properties of SUMO-binding proteins in arabidopsis",

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 detected 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 SUMO1ΔGG or AtSUMO3. The results suggest specific interactions between sumoylation targets and particular sumoylation enzymes. {\textcopyright}2011 KSMCB.",

author = "Hyeongcheol Park and Wonkyun Choi and Heejin Park and Misun Cheong and Yoonduck Koo and Gilok Shin and Woosik Chung and Woeyeon Kim and Mingab Kim and Bressan, {Ray Anthony} and Bohnert, {Hans J{\"u}rgen} and Sangyeol Lee and Daejin Yun",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: We thank Dr. Jian-Min Zhou for providing the NLuc and CLuc plasmids and Dr. Katsunori Tanaka for providing the pCDFDuet-AtSUMO-AtSCE1, pACYCDuet-AtSAEb-AtSAE2, and pET28a-AtMYB30 plasmids. This work was supported by grants from the Biogreen 21 Program (grant No. PJ006654) of the Rural Development Administration, World Class University Program (R32-10148) funded by the Ministry of Education, Science and Technology in Korea, and the National Research Foundation of Korea Grant funded by the Korean Government (Ministry of Education, Science and Technology) [NRF-2010-359-F00006]. GS was supported by scholarship from the Brain Korea 21 program of the Ministry of Education, Science and Technology in Korea.",

year = "2011",

month = may,

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doi = "10.1007/s10059-011-2297-3",

language = "English (US)",

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pages = "143--151",

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AU - Park, Hyeongcheol

AU - Choi, Wonkyun

AU - Park, Heejin

AU - Cheong, Misun

AU - Koo, Yoonduck

AU - Shin, Gilok

AU - Chung, Woosik

AU - Kim, Woeyeon

AU - Kim, Mingab

AU - Bressan, Ray Anthony

AU - Bohnert, Hans Jürgen

AU - Lee, Sangyeol

AU - Yun, Daejin

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: We thank Dr. Jian-Min Zhou for providing the NLuc and CLuc plasmids and Dr. Katsunori Tanaka for providing the pCDFDuet-AtSUMO-AtSCE1, pACYCDuet-AtSAEb-AtSAE2, and pET28a-AtMYB30 plasmids. This work was supported by grants from the Biogreen 21 Program (grant No. PJ006654) of the Rural Development Administration, World Class University Program (R32-10148) funded by the Ministry of Education, Science and Technology in Korea, and the National Research Foundation of Korea Grant funded by the Korean Government (Ministry of Education, Science and Technology) [NRF-2010-359-F00006]. GS was supported by scholarship from the Brain Korea 21 program of the Ministry of Education, Science and Technology in Korea.

PY - 2011/5/20

Y1 - 2011/5/20

N2 - 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 detected 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 SUMO1ΔGG or AtSUMO3. The results suggest specific interactions between sumoylation targets and particular sumoylation enzymes. ©2011 KSMCB.

AB - 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 detected 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 SUMO1ΔGG or AtSUMO3. The results suggest specific interactions between sumoylation targets and particular sumoylation enzymes. ©2011 KSMCB.

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UR - http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3887670

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U2 - 10.1007/s10059-011-2297-3

DO - 10.1007/s10059-011-2297-3

M3 - Article

C2 - 21607647

SN - 1016-8478

VL - 32

SP - 143

EP - 151

JO - Molecules and Cells

JF - Molecules and Cells

IS - 2

ER -

Identification and molecular properties of SUMO-binding proteins in arabidopsis

Abstract

Bibliographical note

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