(a) DARTS assay suggests that ETT binds IAA. Seedlings from an inducible two-component ETT-FLAG line were used for the protein isolation. Samples were treated with DMSO (mock), IAA and BA and …
Parameters for HSQC NMR.
The red boxes indicate the regions used in Figure 1a.
Expression of the canonical auxin responsive IAA19 gene (a) and the ETT-target genes HEC1 (b) and PID (c) in control-treated (dark grey) or 100 µM IAA-treated (light grey) gynoecia assayed using …
Oligonucleotides used in this study.
Output of statistical tests.
In wild-type gynoecia HEC1 and PID are up-regulated upon auxin treatment while both genes are constitutively up-regulated in ett-3. Treatment with 100 µM IAA does not affect HEC1 and PID expression …
(a) Schematic representation of full-length ETT protein highlighting an EAR-like motif and tryptophan in position 505 (W) in the C-terminal ETT-specific domain. DBD, DNA-binding domain. (b) Y2H …
Output of statistical tests.
(a) Alignment of ETT protein sequences of 22 species identified a conserved repressive motif (RLFGF) at its C-terminal domain. A logo of the sequence surrounding the conserved W505 is also shown. (b)…
(a) In Y2H increasing concentrations of IAA lead to reduction of yeast growth abolishing the interaction between ETT and its partners. The interactions are, therefore, auxin-sensitive. (b) Y2H to …
The red boxes indicate the regions used in Figure 3c,d and Figure 3—figure supplement 2d.
(a–d) Promoter GUS expression analysis of pETT:GUS (a), pTPL:GUS (b), pTPR2:GUS (c) and pTPR4:GUS (d) revealed that ETT, TPL and TPR2 but not TPR4 are co-expressed in the Arabidopsis style. Scale …
Output of statistical tests.
Expression analysis using qRT-PCR in wild-type gynoecia showed that TPL and TPR2 are more strongly expressed than TPR1,3 and 4. Likewise, HDA19 exhibits higher expression compared to HDA6. …
(a–c) Chromatin immunoprecipitation (ChIP) shows ETT (a), TPL (b) and HDA19 (c) binding to conserved regions of HEC1 and PID loci in the absence of IAA (dark grey columns). In the presence of IAA …
Output of statistical tests.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information | ||||
---|---|---|---|---|---|---|---|---|
Gene (Arabidopsis thaliana) | ETTIN (ETT) | The Arabidopsis Information Resource | AT2G33860 | |||||
Gene (Arabidopsis thaliana) | TOPLESS (TPL) | The Arabidopsis Information Resource | AT1G15750 | |||||
Gene (Arabidopsis thaliana) | TOPLESS- RELATED 1 (TPR1) | The Arabidopsis Information Resource | AT1G80490 | |||||
Gene (Arabidopsis thaliana) | TOPLESS-RELATED 2 (TPR2) | The Arabidopsis Information Resource | AT3G16830 | |||||
Gene (Arabidopsis thaliana) | TOPLESS-RELATED 3 (TPR3) | The Arabidopsis Information Resource | AT5G27030 | |||||
Gene (Arabidopsis thaliana) | TOPLESS-RELATED 4 (TPR4) | The Arabidopsis Information Resource | AT3G15880 | |||||
Gene (Arabidopsis thaliana) | HISTONE DEACETYLASE 6 (HDA6) | The Arabidopsis Information Resource | AT5G63110 | |||||
Gene (Arabidopsis thaliana) | HISTONE DEACETYLASE 19 (HDA19) | The Arabidopsis Information Resource | AT4G38130 | |||||
Gene (Arabidopsis thaliana) | HECATE 1 (HEC1) | The Arabidopsis Information Resource | AT5G67060 | |||||
Gene (Arabidopsis thaliana) | PINOID (PID) | The Arabidopsis Information Resource | AT2G34650 | |||||
Genetic reagent Arabidopsis thaliana | Col-0 | widely distributed | ||||||
Genetic reagent Arabidopsis thaliana | hda19-4 | Kim et al., 2008 | SALK_13944 | |||||
Genetic reagent Arabidopsis thaliana | tir1-1 afb2-3 afb3-4 | Parry et al., 2009 | ||||||
Genetic reagent Arabidopsis thaliana | tpl | European Arabidopsis Stock Centre | SALK_034518C | |||||
Genetic reagent Arabidopsis thaliana | ett-3 | Simonini et al., 2017; Sessions et al., 1997 | AT2G33860 | |||||
Genetic reagent Arabidopsis thaliana | pETT:GUS | Ng et al., 2009 | AT2G33860 | |||||
Genetic reagent Arabidopsis thaliana | pETT:ETT-GFP | Simonini et al., 2016 | AT2G33860 | |||||
Genetic reagent Arabidopsis thaliana | pTPL:TPL-GFP | Pi et al., 2015 | AT1G15750 | |||||
Genetic reagent Arabidopsis thaliana | p35S:HDA19-GFP | Pi et al., 2015 | AT4G38130 | |||||
Genetic reagent Arabidopsis thaliana | pTIR1:ccvTIR1 | Szemenyei et al., 2008 | ||||||
Genetic reagent Arabidopsis thaliana | tpl tpr2ge | this paper | AT1G15750, AT3G16830 | Further details in the Materials and methods section | ||||
Genetic reagent Arabidopsis thaliana | pTPL:GUS | this paper | AT1G15750 | Further details in the Materials and methods section | ||||
Genetic reagent Arabidopsis thaliana | pTPR2:GUS | this paper | AT3G16830 | Further details in the Materials and methods section | ||||
Genetic reagent Arabidopsis thaliana | pTPR4:GUS | this paper | AT3G15880 | Further details in the Materials and methods section | ||||
Genetic reagent Arabidopsis thaliana | p35S:CDGVG p6xGAL4UAS:ETT-FLAG | this paper | Further details in the Materials and methods section | |||||
Antibody | Mouse anti-FLAG | Abcam | ab49763 | monoclonal, conjugated with HRP | ||||
Antibody | Mouse anti-HA | Abcam | ab173826 | monoclonal, conjugated with HRP | ||||
Antibody | Mouse anti-GFP | Roche | 11814460001 | monoclonal | ||||
Antibody | Rabbit anti-H3K27ac | Abcam | ab4729 | polyclonal | ||||
Antibody | Rabbit anti-H3 | Abcam | ab1791 | polyclonal | ||||
Recombinant DNA reagent | pGWB14; p35s:HA-TPL | Espinosa-Ruiz et al., 2017 | ||||||
Recombinant DNA reagent | pICH47732; p35s:ETT-3xFLAG | this paper | Backbone Addgene #48000 | |||||
Recombinant DNA reagent | pICH47732; p35s: ETTW505A-3xFLAG | this paper | Backbone Addgene #48000 | |||||
Recombinant DNA reagent | pGADT7; ETT | this paper | Further details in the Materials and methods section | |||||
Recombinant DNA reagent | pGADT7; ETTW505A | this paper | Further details in the Materials and methods section | |||||
Recombinant DNA reagent | pGADT7; ETTL552S; F553S | this paper | Further details in the Materials and methods section | |||||
Recombinant DNA reagent | pGBKT7; TPL | Causier et al., 2012 | ||||||
Recombinant DNA reagent | pGBKT7; TPR1 | Causier et al., 2012 | ||||||
Recombinant DNA reagent | pGBKT7; TPR2 | Causier et al., 2012 | ||||||
Recombinant DNA reagent | pGBKT7; TPR3 | Causier et al., 2012 | ||||||
Recombinant DNA reagent | pGBKT7; TPR4 | Causier et al., 2012 | ||||||
Recombinant DNA reagent | pESPRIT; ES388-594 | Simonini et al., 2018 |