Identification and molecular properties of SUMO-binding proteins in arabidopsis

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.

Park, H. C., Choi, W., Park, H. J., Cheong, M. S., Koo, Y. D., Shin, G., … Yun, D.-J. (2011). Identification and molecular properties of SUMO-binding proteins in Arabidopsis. Molecules and Cells, 32(2), 143–151. doi:10.1007/s10059-011-2297-3

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.

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