Journal Article

Fluorescence Cross-Correlation Analyses of the Molecular Interaction between an Aux/IAA Protein, MSG2/IAA19, and Protein–Protein Interaction Domains of Auxin Response Factors of Arabidopsis Expressed in HeLa Cells

Hideki Muto, Issei Nagao, Taku Demura, Hiroo Fukuda, Masataka Kinjo and Kotaro T. Yamamoto

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 47, issue 8, pages 1095-1101
Published in print August 2006 | ISSN: 0032-0781
Published online August 2006 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcj080
Fluorescence Cross-Correlation Analyses of the Molecular Interaction between an Aux/IAA Protein, MSG2/IAA19, and Protein–Protein Interaction Domains of Auxin Response Factors of Arabidopsis Expressed in HeLa Cells

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Since auxin may elicit numerous developmental responses by the use of a combination of auxin response factors (ARFs) and their Aux/IAA repressors, it is important to determine the interaction between the two protein families in a quantitative manner. We transiently expressed the C-terminal protein–protein interaction domains (CTDs) of Arabidopsis ARFs, MP/ARF5 and NPH4/ARF7, and MSG2/IAA19, fused to fluorescent proteins in HeLa cells, and determined their molecular interactions with fluorescence cross-correlation spectroscopy (FCCS). Almost complete association was found between MSG2 and MP-CTD and between MSG2 and NPH4-CTD. Approximately 20% association was found for MSG2 homodimers, NPH4-CTD homodimers and MP-CTD/NPH4-CTD heterodimers. Homotypic binding of MP-CTD may be weaker than that of MSG2. MSG2 was localized in cytoplasmic compartments in HeLa cells, whereas it was localized in the nuclei in plant cells. The fact that the heterotypic interaction between MSG2 and ARF-CTDs is stronger than each of the homotypic interactions appears to be the molecular basis for tight control of the transcriptional activity of ARFs by auxin. These results also show that FCCS is useful to examine protein–protein interactions especially for transcriptional regulators.

Keywords: Arabidopsis thaliana — Aux/IAA protein — Auxin response factor — Fluorescence cross-correlation spectroscopy — HeLa cell — Protein–protein interaction

Journal Article.  3755 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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