Minvenus Bio - A Higginsianin-Independent Regulator of JAZ Protein Degradation

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Affecting JAZ protein degradation

To investigate the effect on higginsianins of JA-induced degradation in vivo of the JAIle sensor, we used Arabidopsis seeds constitutively expressing a JA degron domain fused with VENUS yellow fluorescence protein and a nucleolar localization signal (Jas9VENUS, Larrieu et. al., 2015). Prior to each treatment, the roots of these seedlings were pre-treated for 30 min with either mock or compounds under analysis. Treatment of the seedlings with MeJA induced J9-VENUS reporter degradation, as shown by decreased fluorescence intensity in nuclei of the cells (Figure 3). Higginsianin B pre-treatment of the seedlings significantly inhibited MeJA-induced degradation.

Venus-Hkb was expressed in cells at the site of an invagination hole forming as early as t=63 min prior to invagination. The expression increased and covered a large area across the platcode (Figure 5 figure supplement 1B-C). In contrast, Fkh-GFP was expressed close to the placode but decreased with a delay, suggesting that Hkb and Fkh are regulated differently by JA.

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