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Small‐molecule auxin inhibitors that target YUCCA are powerful tools for studying auxin function
Authors:Yusuke Kakei  Chiaki Yamazaki  Masashi Suzuki  Ayako Nakamura  Akiko Sato  Yosuke Ishida  Rie Kikuchi  Shouichi Higashi  Yumiko Kokudo  Takahiro Ishii  Kazuo Soeno  Yukihisa Shimada
Affiliation:1. Kihara Institute for Biological Research, Yokohama City University, Yokohama, Kanagawa, Japan;2. Graduate School of Nanobioscience, Yokohama City University, Yokohama, Japan;3. National Agriculture and Food Research Organization (NARO), Western Region Agricultural Research Center (WARC), Zentsuji, Kagawa, Japan
Abstract:Auxin is essential for plant growth and development, this makes it difficult to study the biological function of auxin using auxin‐deficient mutants. Chemical genetics have the potential to overcome this difficulty by temporally reducing the auxin function using inhibitors. Recently, the indole‐3‐pyruvate (IPyA) pathway was suggested to be a major biosynthesis pathway in Arabidopsis thaliana L. for indole‐3‐acetic acid (IAA), the most common member of the auxin family. In this pathway, YUCCA, a flavin‐containing monooxygenase (YUC), catalyzes the last step of conversion from IPyA to IAA. In this study, we screened effective inhibitors, 4‐biphenylboronic acid (BBo) and 4‐phenoxyphenylboronic acid (PPBo), which target YUC. These compounds inhibited the activity of recombinant YUC in vitro, reduced endogenous IAA content, and inhibited primary root elongation and lateral root formation in wild‐type Arabidopsis seedlings. Co‐treatment with IAA reduced the inhibitory effects. Kinetic studies of BBo and PPBo showed that they are competitive inhibitors of the substrate IPyA. Inhibition constants (Ki) of BBo and PPBo were 67 and 56 nm , respectively. In addition, PPBo did not interfere with the auxin response of auxin‐marker genes when it was co‐treated with IAA, suggesting that PPBo is not an inhibitor of auxin sensing or signaling. We propose that these compounds are a class of auxin biosynthesis inhibitors that target YUC. These small molecules are powerful tools for the chemical genetic analysis of auxin function.
Keywords:auxin biosynthesis inhibitor  chemical biology     YUCCA        Arabidopsis thaliana     borate     Brachypodium distachyon     technical advance
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