BLADE-ON-PETIOLE proteins act in an E3 ubiquitin ligase complex to regulate PHYTOCHROME INTERACTING FACTOR 4 abundance
- Swedish University of Agricultural Sciences, Sweden
- University of Lausanne, Switzerland
- Umeå University, Sweden
Figures
Figure 1 with 3 supplements
BOP2 promotes photomorphogenesis in red light.
(a) Hypocotyl lengths of 5-day-old Col-0, bop1, bop2, and bop1 bop2 seedlings grown in constant red light (Rc) of different fluence rates with phyB mutants included as controls. (b) Hypocotyl …
Figure 1—figure supplement 1
Hypocotyl lengths and hook angles of BOP mutants in response to white light and monochromatic light.
(a–c) Hypocotyl lengths of 5-day-old Col-0, bop1, bop2, and bop1 bop2 seedlings grown in constant white light (a), blue light (b), and far-red light (c) of different fluence rates, using cry1, cry2, …
Figure 1—figure supplement 2
Hypocotyl lengths, cotyledon angles, and hook angles of BOP1 and BOP2 overexpressing lines in response to red light.
(a) Hypocotyl lengths of Col-0, 35S::BOP1 and 35S::BOP2 seedlings grown in constant red light (Rc) of different fluence rates with phyB mutant included as controls. Lower panel shows the hypocotyl …
Figure 1—figure supplement 3
bop2-2 shows wild-type accumulation of phyB levels in 5 day-old seedlings grown in constant red light.
(a) Hypocotyl lengths of Col-0, phyB and phyB bop2 seedlings grown at constant red light (Rc) of different fluence rates. Right panel shows the hypocotyl length phenotype of indicated lines. (b) …
Figure 2 with 1 supplement
BOP2 genetically interacts with PIF4 in response to red light.
(a) Hypocotyl lengths of 5-day-old Col-0, bop2, pif4, and pif4 bop2 seedlings grown in constant red light (Rc) of different fluence rates. (b) Hypocotyl phenotypes of indicated seedlings in dark and …
Figure 2—figure supplement 1
Kinematic analysis of hypocotyl lengths.
Kinematic analysis of hypocotyl lengths in response to red light for Col-0, pif4, bop1 bop2, and pif4 bop1 bop2 seedlings. Seedlings were firstly grown in dark for 3 days then switch to 6 μmol·m−2·s−…
Figure 3 with 1 supplement
BOP2 promotes degradation of PIF4 in response to red light.
(a) Hypocotyl lengths of 5-day-old Col-0, pif4, pif4 bop2, pif4;PIF4p::PIF4-HA, and pif4 bop2;PIF4p::PIF4-HA seedlings grown in constant red light (Rc) of different fluence rates. The data are shown …
Figure 3—figure supplement 1
Hypocotyl lengths of 35S::PIF4-HA and 35S::PIF4-HA bop2 plants.
Hypocotyl lengths of 5-day-old Col-0, bop2, 35S::PIF4-HA, and 35S::PIF4-HA bop2 seedlings grown in constant red light (Rc) of different fluence rates. Quantitative data are shown as means ± s.d., n =…
Figure 4 with 1 supplement
BOP1 and BOP2 interact with CUL3 and PIF4.
(a) Co-immunoprecipitation assays of CUIL3A and BOP1/2 interaction in Arabidopsis protoplasts. Co-immunoprecipitated HA-CUL3A was detected by western blotting using anti-HA antibodies, after …
Figure 4—figure supplement 1
Protein-protein interaction analysis.
(a) Co-immunoprecipitation analysis of CUL3A and BOP2 interaction in a 35S::myc-BOP2 transgenic line. Co-immunoprecipitated CUL3A was detected by western blotting using anti-CUL3A antibodies (Enzo …
Figure 5 with 1 supplement
CUL3-BOP complex mediates the polyubiquitination of PIF4.
(a) TUBEs (tandem ubiquitin binding entities) assays of ubiquitinated proteins in dark and red light treated pif4;PIF4p::PIF4-HA and pif4bop2;PIF4p::PIF4-HA seedlings. 3-day-old dark-grown seedlings …
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Figure 5—source data 1
Quantification of ubiquitinated PIF4-HA protein levels relative to total ubiquitinated proteins from TUBEs assays.
- https://doi.org/10.7554/eLife.26759.014
Figure 5—figure supplement 1
BOP2 mediates ubiquitination of PIF4-HA also in darkness.
3-day-old dark-grown pif4;PIF4p::PIF4-HA and pif4bop2;PIF4p::PIF4-HA seedlings were irradiated with 6 μmol·m−2·s−1 red light for 2 min followed by 8 min in the dark before harvesting. Total …
Figure 6 with 2 supplements
BOP2 modulates the PIF4 abundance in temperature response.
(a) Hypocotyl lengths of indicated seedlings in response to the temperature of 28°C. Seedlings were grown firstly at 22°C in constant white light for 4 days then transferred to 28°C or kept in 22°C …
Figure 6—figure supplement 1
Hypocotyl lengths of indicated seedlings in response to different temperatures.
Hypocotyl lengths of indicated seedlings in response to the temperature of 28°C in 12 hr light/12 hr dark condition (a) and in constant white light (b). Seedlings were grown firstly at 22°C for 4 …
Figure 6—figure supplement 2
Genetic interactions between bop1, bop2 and phyB.
Plants were grown for 20 days in constant red light. The two arrow heads delimit one rosette internode.
Additional files
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Supplementary file 1
All primer sequences used in this study.
- https://doi.org/10.7554/eLife.26759.018
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Transparent reporting form
- https://doi.org/10.7554/eLife.26759.019
