In vitro and in vivo interaction of AtRma2 E3 ubiquitin ligase and auxin binding protein 1

Ora Son, Seok Keun Cho, Soo Jin Kim, Woo Taek Kim

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

E3 ubiquitin (Ub) ligases play diverse roles in cellular regulation in eukaryotes. Three homologous AtRmas (AtRma1, AtRma2, and AtRma3) were recently identified as ER-localized Arabidopsis homologs of human RING membrane-anchor E3 Ub ligase. Here, auxin binding protein 1 (ABP1), one of the auxin receptors in Arabidopsis, was identified as a potential substrate of AtRma2 through a yeast two-hybrid assay. An in vitro pull-down assay confirmed the interaction of full-length AtRma2 with ABP1. AtRma2 was transiently expressed in tobacco (Nicotiana benthamiana) plants through an Agrobacterium-mediated infiltration method and bound ABP1 in vivo. In vitro ubiquitination assays revealed that bacterially-expressed AtRma2 ubiquitinated ABP1. ABP1 was poly-ubiquitinated in tobacco cells and its stability was significantly increased in the presence of MG132, a 26S proteasome inhibitor. This suggests that ABP1 is controlled by the Ub/26S proteasome system. Therefore, AtRma2 is likely involved in the cellular regulation of ABP1 expression levels.

Original languageEnglish
Pages (from-to)492-497
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume393
Issue number3
DOIs
Publication statusPublished - 2010 Mar 12

Fingerprint

Ubiquitin-Protein Ligases
Tobacco
Assays
Arabidopsis
Agrobacterium
Two-Hybrid System Techniques
Proteasome Inhibitors
Indoleacetic Acids
Ubiquitination
Ubiquitin
auxin-binding protein 1
In Vitro Techniques
Eukaryota
Anchors
Infiltration
Yeast
Membranes
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

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In vitro and in vivo interaction of AtRma2 E3 ubiquitin ligase and auxin binding protein 1. / Son, Ora; Cho, Seok Keun; Kim, Soo Jin; Kim, Woo Taek.

In: Biochemical and Biophysical Research Communications, Vol. 393, No. 3, 12.03.2010, p. 492-497.

Research output: Contribution to journalArticle

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