Cullin, a component of the SCF complex, interacts with TaRMD5 during wheat spike development

M. J. Hong, D. Y. Kim, Y. W. Seo

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    4 Citations (Scopus)

    Abstract

    Cullin, a major component of the SKP1-cullin-F box protein (SCF) complex, is a scaffold protein that binds to both SKP1 and RBX1 for selective protein degradation through the ubiquitin proteasome system. In order to study the role of cullin in common wheat, we isolated TaCullin (Cullin gene from Triticum aestivum) from wheat spike cDNA. TaCullin was expressed during all spike/grain developmental stages and in high amounts during early spike/grain development. The TaCullin gene is located on the chromosome arm 2DL. Our results suggest that unneddylated TaCullin is located in the nucleus. Based on previous proposals of Cullin-SKP1 interactions, we examined the interaction between TaCullin and SKP1-like protein (TaSKP) families by using a yeast two-hybrid approach. Yeast cotransformation demonstrated that the N-terminus of TaCullin physically interacts with TaSKP proteins. Using the yeast two-hybrid screen, we identified potential TaCullin-interacting proteins in a wheat spike library. Among the 9 clones that were identified as potential interacting partners of TaCullin, we identified E3-like ubiquitin ligase, targeting fructose-1,6-bisphosphatase (RMD5) homolog A-like protein. The interaction between TaCullin and the TaRMD5 homolog A-like protein was specifically mediated through the C-terminus of TaCullin. The results of bimolecular fluorescence complementation assay indicated that TaCullin-TaRMD5 is localized in the plasma membrane and cytoplasm. In this study, we present that TaRMD5, such as RING box protein 1 (RBX1), has the potential to interact with TaCullin, depending on the developmental stage and particular organ tissues analyzed.

    Original languageEnglish
    Pages (from-to)218-230
    Number of pages13
    JournalBiologia Plantarum
    Volume58
    Issue number2
    DOIs
    Publication statusPublished - 2014 Jun

    Bibliographical note

    Funding Information:
    Submitted 10 January 2013, last revision 11 July 2013, accepted 15 July 2013. Abbreviations: APC/C - anaphase-promoting complex or cyclosome; BiFC - bimolecular fluorescence complementation assay; CAND1 - cullin-associated and neddylation-dissociated 1; CRL - cullin-RING ubiquitin ligases; DAF - day after flowering; GFP - green fluorescent protein; HECT - homologous to the E6-AP carboxyl terminus; ONPG - o-nitrophenyl-β-galactoside; RBX1 - RING box protein 1; RING - really interesting new gene; RMD5 - E3-like ubiquitin ligase targeting fructose-1,6-bisphosphatase; SCF - SKP1-cullin-F box protein; SKP - S-phase kinase-associated protein 1; UPS - ubiquitin proteasome system; Y2H - yeast two-hybrid. Acknowledgements: This research was supported by the Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea, by a grant from the Next-Generation Bio-Green 21 Program (Plant Molecular Breeding Center No. PJ0080312012), and by the National Nuclear R&D Program through the National Research Foundation of Korea (No. 2012M2A2A60-35566). * Corresponding author; fax: (+82) 2 3290-3501, e-mail: [email protected]

    Keywords

    • TaSKP
    • Triicum aestivum
    • chromosome location
    • yeast two-hybrid screening

    ASJC Scopus subject areas

    • Plant Science
    • Horticulture

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