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Abstract Photoactivated phytochrome B (PHYB) binds to antagonistically acting PHYTOCHROME-INTERACTING transcription FACTORs (PIFs) to regulate hundreds of light responsive genes in Arabidopsis by promoting PIF degradation. However, whether PHYB directly controls the transactivation activity of PIFs remains ambiguous. Here we show that the prototypic PIF, PIF3, possesses a p53-like transcription activation domain (AD) consisting of a hydrophobic activator motif flanked by acidic residues. A PIF3mAD mutant, in which the activator motif is replaced with alanines, fails to activate PIF3 target genes in Arabidopsis , validating the functions of the PIF3 AD in vivo. Intriguingly, the N-terminal photosensory module of PHYB binds immediately adjacent to the PIF3 AD to repress PIF3’s transactivation activity, demonstrating a novel PHYB signaling mechanism through direct interference of the transactivation activity of PIF3. Our findings indicate that PHYB, likely also PHYA, controls the stability and activity of PIFs via structurally separable dual signaling mechanisms.
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This content will become publicly available on June 17, 2026
Distinct signaling mechanisms of constitutively active phytochromes A and B in regulating Arabidopsis seedling development
Plant phytochromes are well-studied photoreceptors that sense red and far-red light, regulating photomorpho- genic development. Molecular signaling mechanisms of phytochrome A (phyA) and phyB largely overlap, especially in regulation of PHYTOCHROME-INTERACTING FACTORs (PIFs) and E3 ligase complexes composed of CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) and SUPPRESSORs OF phyA-105 (SPAs). However, the differences in their molecular signaling mechanisms remain unclear. Constitutively active mutants of phyB (YVB) and NLS-fused phyA (YVA:NLS) mediate light-independent seedling development, leading to constitutive photomorphogenic (cop) phenotypes in their transgenic Arabidopsis plants. Interestingly, YVB interacted with PIF3 independently of light, but YVA showed little interaction. In this study, we investigated distinct signaling mechanisms underlying the similar cop phenotypes given by YVB and YVA:NLS. Our findings indicated that YVA efficiently inactivate the COP1/SPA complex, leading to accumulation of ELONGATED HYPOCOTYL 5 (HY5) and subsequent expression of its target genes HY5 and HYH. YVB induced light-independent PIF3 and PIF1 degra- dation, in addition to HY5 accumulation. Moreover, co-expression of PIF3 in the YVB plant significantly attenuated the cop phenotypes, but minimal effects were observed in the YVA:NLS plant. In particular, PIF3 negatively regulated the interaction between YVB and COP1, which decreased HY5 accumulation in the YVB plant co-expressing PIF3. Furthermore, when transferred from light to dark, PIF3 was highly accumulated in phyB-5, whereas HY5 is degraded faster in phyA-201 compared to that in Ler. Collectively, our results suggest HY5 accumulation as the molecular bases for the cop phenotypes and also indicate that phyB is more important for regulating PIF3, whereas phyA effectively inactivates the COP1/SPA complex relative to PIF3 degradation.
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- Award ID(s):
- 2014408
- PAR ID:
- 10639132
- Publisher / Repository:
- www.elsevier.com
- Date Published:
- Journal Name:
- Plant physiology and biochemistry
- Volume:
- 229
- ISSN:
- 1873-2690
- Page Range / eLocation ID:
- 110412
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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