Li S., Gutsche N., and Zachgo S. The ROXY1 C-Terminal L**LL Motif Is Essential for the Interaction with TGA Transcription Factors.
In Figure 3A, one bimolecular fluorescence complementation picture shows the typical ROXY1/TGA nuclear localization observed for YN-ROXY1/YC-TGA3 (reported before by Li et al. [2009], Plant Cell) and YN-ROXY1Ɗ4/YC-TGA3. A nuclear interaction was also observed for YN-ROXY1/YC-PAN proteins, as indicated in the legend and reported before by Li et al. (2009). Figure 3A shows the ROXY1/PAN interaction that was taken from the Plant Cell 2009 publication (figure 2, top left) without proper indication. Bimolecular fluorescence complementation analyses of the three protein interactions and also for YN-ROXY1Ɗ4/PAN have been repeated, confirming nuclear interactions for all tested protein pairs. The data were submitted to the journal for inspection during the correction process.
In planta interactions of mutagenized ROXY1 with TGA proteins and subcellular localization of ROXY1 variants. A, Wild-type ROXY1 and the truncated form of ROXY1 lacking the C-terminal four amino acids (ROXY1Ɗ4) interact with TGA3 in the nucleus. The same nuclear interaction was also observed for PAN. B, All investigated ROXY1 substitution variants (ROXY1L1A, ROXY1L2A, ROXY1L3A, ROXY1L2/3A, and ROXY1L1/2/3A) failed to interact in planta, which is shown exemplarily for ROXY1L1A. C, Nucleocytoplasmic distribution of the GFP/ROXY1 fusion protein. D, Replacement of the first Leu (L1) with an Ala in the ROXY1 L**LL and removal of the ROXY1 C-terminal four amino acids showed a wild-type-like nucleocytoplasmic localization. All other mutagenized ROXY1 proteins also accumulated in the nucleus and the cytoplasm. Bars = 50 μm. The image shown in A shows a typical nuclear interaction of ROXY1 with TGA proteins. The image was duplicated from figure 2 (top left) fromLi et al. (2009), which shows a ROXY1/PAN interaction. Additional information onFigure 3Aalso is shown.
The abnormal petal phenotype of the Arabidopsis (Arabidopsis thaliana) roxy1 mutant and the potential of ROXYs to complement the roxy1 flower phenotype have been described in several other publications (Xing et al., 2005; Li et al., 2009; Wang et al., 2009; Gutsche et al., 2017). The formation of flowers with four versus two or five petals was used as a qualitative trait marker for the categorization of flowers into wild-type-like versus roxy1-like flower phenotypes. The four images in Figure 4 show representative wild-type and roxy1 flower phenotypes, as stated in the legend text. Images were taken from figure 3D and figure 1B from the Plant Cell 2009 article without proper indication. The essence of the presented findings is correct, as the shown images represent the key qualitative differences in petal morphology between the different studied genotypes.
Complementation experiments of mutagenized ROXY1 proteins. Wild-type and mutant ROXY genes driven by the ROXY1 promoter were transformed into the roxy1 mutant. Petal phenotypes of transgenic T1 plants were scored, and representative phenotypes are shown. The roxy1 mutant flowers (B) develop only 2.5 petals per flower instead of 4.0 petals as observed in wild-type (WT) flowers (A). The wild-type ROXY1 gene is able to complement the roxy1 mutant (C). In contrast, replacement of the first Leu (L1) with an Ala in the L**LL motif of ROXY1 (ROXY1L1A) or removal of the ALWL motif from the ROXY1 C terminus (ROXY1Ɗ4) failed to rescue abnormal petal phenotypes of the roxy1 mutant (D). All other mutagenized ROXY1 proteins (ROXY1L2A, ROXY1L3A, ROXY1L2,3A, and ROXY1L1,2,3A) also lost their capacities to complement the roxy1 mutant. At least 50 T1 transformants were analyzed for each construct. The images shown inFigure 4were duplicated fromLi et al. (2009). Figure 4Awas duplicated from figure 3D (top right),Figure 4Bfrom figure 3D (bottom right),Figure 4Cfrom figure 3D (top left), andFigure 4Dfrom figure 1B (bottom left, flipped horizontally).
The legends have been amended accordingly with added text shown in bold. The errors described in these figures do not affect the conclusions of this article.
LITERATURE CITED
