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Volume 23
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Article

Fluorescent Auxin Analogs Report Two Auxin Binding Sites with Different Subcellular Distribution and Affinities: A Cue for Non-Transcriptional Auxin Signaling

by
Xiang Huang
1,2,
Jan Maisch
1,
Ken-Ichiro Hayashi
3 and
Peter Nick
1,*
1
Molecular Cell Biology, Botanical Institute, Karlsruhe Institute of Technology, Fritz-Haber-Weg 4, 76133 Karlsruhe, Germany
2
Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510301, China
3
Department of Biochemistry, Okayama University of Science, 1-1 Ridai-cho, Okayama 700-0005, Japan
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2022, 23(15), 8593; https://doi.org/10.3390/ijms23158593
Submission received: 30 June 2022 / Revised: 1 August 2022 / Accepted: 1 August 2022 / Published: 2 August 2022
(This article belongs to the Section Molecular Plant Sciences)
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Abstract

The complexity of auxin signaling is partially due to multiple auxin receptors that trigger differential signaling. To obtain insight into the subcellular localization of auxin-binding sites, we used fluorescent auxin analogs that can undergo transport but do not deploy auxin signaling. Using fluorescent probes for different subcellular compartments, we can show that the fluorescent analog of 1-naphthaleneacetic acid (NAA) associates with the endoplasmic reticulum (ER) and tonoplast, while the fluorescent analog of indole acetic acid (IAA) binds to the ER. The binding of the fluorescent NAA analog to the ER can be outcompeted by unlabeled NAA, which allows us to estimate the affinity of NAA for this binding site to be around 1 μM. The non-transportable auxin 2,4-dichlorophenoxyacetic acid (2,4-D) interferes with the binding site for the fluorescent NAA analog at the tonoplast but not with the binding site for the fluorescent IAA analog at the ER. We integrate these data into a working model, where the tonoplast hosts a binding site with a high affinity for 2,4-D, while the ER hosts a binding site with high affinity for NAA. Thus, the differential subcellular localization of binding sites reflects the differential signaling in response to these artificial auxins.
Keywords: auxin; indole acetic acid; binding site; NBD auxins; ER; tonoplast; auxin signaling auxin; indole acetic acid; binding site; NBD auxins; ER; tonoplast; auxin signaling

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MDPI and ACS Style

Huang, X.; Maisch, J.; Hayashi, K.-I.; Nick, P. Fluorescent Auxin Analogs Report Two Auxin Binding Sites with Different Subcellular Distribution and Affinities: A Cue for Non-Transcriptional Auxin Signaling. Int. J. Mol. Sci. 2022, 23, 8593. https://doi.org/10.3390/ijms23158593

AMA Style

Huang X, Maisch J, Hayashi K-I, Nick P. Fluorescent Auxin Analogs Report Two Auxin Binding Sites with Different Subcellular Distribution and Affinities: A Cue for Non-Transcriptional Auxin Signaling. International Journal of Molecular Sciences. 2022; 23(15):8593. https://doi.org/10.3390/ijms23158593

Chicago/Turabian Style

Huang, Xiang, Jan Maisch, Ken-Ichiro Hayashi, and Peter Nick. 2022. "Fluorescent Auxin Analogs Report Two Auxin Binding Sites with Different Subcellular Distribution and Affinities: A Cue for Non-Transcriptional Auxin Signaling" International Journal of Molecular Sciences 23, no. 15: 8593. https://doi.org/10.3390/ijms23158593

APA Style

Huang, X., Maisch, J., Hayashi, K.-I., & Nick, P. (2022). Fluorescent Auxin Analogs Report Two Auxin Binding Sites with Different Subcellular Distribution and Affinities: A Cue for Non-Transcriptional Auxin Signaling. International Journal of Molecular Sciences, 23(15), 8593. https://doi.org/10.3390/ijms23158593

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