Evolutionary comparison of competitive protein-complex formation of MYB, bHLH, and WDR proteins in plants

Abstract A protein complex consisting of a MYB, basic Helix-Loop-Helix, and a WDR protein, the MBW complex, regulates five traits, namely the production of anthocyanidin, proanthocyanidin, and seed-coat mucilage, and the development of trichomes and root hairs. For complexes involved in trichome and root hair development it has been shown that the interaction of two MBW proteins can be counteracted by the respective third protein (called competitive complex formation). We examined competitive complex formation for selected MBW proteins from Arabidopsis thaliana, Arabis alpina, Gossypium hirsutum, Petunia hybrida, and Zea mays. Quantitative analyses of the competitive binding of MYBs and WDRs to bHLHs were done by pull-down assays using ProtA- and luciferase-tagged proteins expressed in human HEC cells. We found that some bHLHs show competitive complex formation whilst others do not. Competitive complex formation strongly correlated with a phylogenetic tree constructed with the bHLH proteins under investigation, suggesting a functional relevance. We demonstrate that this different behavior can be explained by changes in one amino acid and that this position is functionally relevant in trichome development but not in anthocyanidin regulation.

Download Full-text

Conserved regulatory mechanism controls the development of cells with rooting functions in land plants

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1416324112 ◽

2015 ◽

Vol 112 (29) ◽

pp. E3959-E3968 ◽

Cited By ~ 43

Author(s):

Thomas Ho Yuen Tam ◽

Bruno Catarino ◽

Liam Dolan

Keyword(s):

Physcomitrella Patens ◽

Lotus Japonicus ◽

Root Hairs ◽

Land Plants ◽

Last Common Ancestor ◽

Core Network ◽

Helix Loop Helix ◽

Root Hair Development ◽

Bhlh Transcription Factors ◽

Bhlh Proteins

Land plants develop filamentous cells—root hairs, rhizoids, and caulonemata—at the interface with the soil. Members of the group XI basic helix–loop–helix (bHLH) transcription factors encoded by LOTUS JAPONICUS ROOTHAIRLESS1-LIKE (LRL) genes positively regulate the development of root hairs in the angiosperms Lotus japonicus, Arabidopsis thaliana, and rice (Oryza sativa). Here we show that auxin promotes rhizoid and caulonema development by positively regulating the expression of PpLRL1 and PpLRL2, the two LRL genes in the Physcomitrella patens genome. Although the group VIII bHLH proteins, AtROOT HAIR DEFECTIVE6 and AtROOT HAIR DEFECTIVE SIX-LIKE1, promote root-hair development by positively regulating the expression of AtLRL3 in A. thaliana, LRL genes promote rhizoid development independently of PpROOT HAIR DEFECTIVE SIX-LIKE1 and PpROOT HAIR DEFECITVE SIX-LIKE2 (PpRSL1 and PpRSL2) gene function in P. patens. Together, these data demonstrate that both LRL and RSL genes are components of an ancient auxin-regulated gene network that controls the development of tip-growing cells with rooting functions among most extant land plants. Although this network has diverged in the moss and the angiosperm lineages, our data demonstrate that the core network acted in the last common ancestor of the mosses and angiosperms that existed sometime before 420 million years ago.

Download Full-text

Faculty Opinions recommendation of Specific and stable fluorescence labeling of histidine-tagged proteins for dissecting multi-protein complex formation.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1031009.364463 ◽

2006 ◽

Author(s):

Wen-hong Li

Keyword(s):

Complex Formation ◽

Protein Complex ◽

Fluorescence Labeling ◽

Protein Complex Formation

Download Full-text

Faculty Opinions recommendation of Conformational heterogeneity in antibody-protein antigen recognition: implications for high affinity protein complex formation.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718241924.793489921 ◽

2014 ◽

Author(s):

Jia-Huai Wang

Keyword(s):

Complex Formation ◽

Protein Complex ◽

Antigen Recognition ◽

Protein Antigen ◽

Conformational Heterogeneity ◽

High Affinity ◽

Protein Complex Formation

Download Full-text

A Pivotal Role of the Basic Leucine Zipper Transcription Factor bZIP53 in the Regulation of Arabidopsis Seed Maturation Gene Expression Based on Heterodimerization and Protein Complex Formation

The Plant Cell ◽

10.1105/tpc.108.062968 ◽

2009 ◽

Vol 21 (6) ◽

pp. 1747-1761 ◽

Cited By ~ 109

Author(s):

Rosario Alonso ◽

Luis Oñate-Sánchez ◽

Fridtjof Weltmeier ◽

Andrea Ehlert ◽

Isabel Diaz ◽

...

Keyword(s):

Gene Expression ◽

Transcription Factor ◽

Complex Formation ◽

Protein Complex ◽

Leucine Zipper ◽

Pivotal Role ◽

Basic Leucine Zipper ◽

Arabidopsis Seed ◽

Protein Complex Formation

Download Full-text

Analysis of Protein Complex Formation on Membrane Surfaces by Single Molecule Diffusion

Biophysical Journal ◽

10.1016/j.bpj.2011.11.994 ◽

2012 ◽

Vol 102 (3) ◽

pp. 183a

Author(s):

Brian P. Ziemba ◽

Jefferson D. Knight ◽

Joseph J. Falke

Keyword(s):

Complex Formation ◽

Single Molecule ◽

Protein Complex ◽

Membrane Surfaces ◽

Protein Complex Formation

Download Full-text

Targeting WNT10B-Driven Signalling through Induction of FZD6 By Porcupine Inhibition in T Cell Acute Lymphoblastic Leukemia

Blood ◽

10.1182/blood-2019-124871 ◽

2019 ◽

Vol 134 (Supplement_1) ◽

pp. 3956-3956

Author(s):

Adriana Cassaro ◽

Francesca Lazzaroni ◽

Giovanni Grillo ◽

Gianluigi Reda ◽

Roberto Cairoli ◽

...

Keyword(s):

Acute Lymphoblastic Leukemia ◽

T Cell ◽

Complex Formation ◽

Wnt Signaling ◽

Protein Complex ◽

Lymphoblastic Leukemia ◽

Protein Complex Formation ◽

Cell Acute Lymphoblastic Leukemia ◽

Short Hairpin ◽

Signaling Activation

Background Wnt/Fzd signaling is known to play a pervasive influence in hematopoietic stem cell maintenance, T-cell development in the thymus and function as well as an important role in T-cell acute lymphoblastic leukemia (T-ALL) establishment. We have previously described a recurrent rearrangement involving the WNT10Blocus (WNT10BR) expressing a transcript variant (WNT10BIVS1) in acute myeloid leukemia. To determine the occurrence of this rearrangement in T-ALL we analyzed retrospectively an italian cohort of patients (n=20) and detected the WNT10BRrearrangement with a high prevalence (14/20). We also confirmed the relevance of these findings to human disease, detecting the molecular circuit triggered by the WNT10B over-expression using the MOLT-4 T-ALL cell model.In this report, we examined the expression of components of the Wnt signaling cascade mediated by WNT10B and the effects of specific gene silencing by short hairpin RNA (shRNA) and exposure to the potent PORCN inhibitor (LGK974), or the TGFbRI inhibitor (A83-01) on the WNT10B-mediated Wnt signaling activation. Methods We used the T-ALL model MOLT-4 cell line to assess the WNT10B/FZD signaling axis driven by WNT10BR. In order to identify interaction between WNT10B and FZD receptors we performed in situ proximity ligation assay (PLA) a method used to visualize protein-protein interactions.MOLT4 cells were infected with WNT10B/WNT10BIVS1-shRNA silencing lentiviral vectors versus empty vector control and treated with increased concentration of LGK974 or A83-01, subsequently the effects of pharmacological inhibition on the WNT10B/FZD interactions and on Wnt effector proteins were evaluated by PLA and expression analyses. Cell proliferation and cell death were measured by EdU assay and Annexin-V/Propidium Iodide (PI) analyses. Results We found that WNT10BRdrives Wnt signaling activity in T-ALL through interaction of WNT10B with FZD6 receptor. The effects of WNT10B/FZD6 interaction on Wnt-mediated signal in MOLT-4 were interfered by short hairpin RNAs (shRNAs)-mediated gene silencing and by small molecules-mediated disruption of Wnt-dependent signaling. We performed WNT10BIVS1knockdown or pharmacological inhibition of WNT10B release by the porcupine (PORCN) inhibitor LGK974 and these in turn progressively down-modulate WNT10B/FZD6 protein complex formation and significantly impairs intracellular effectors and leukemic expansion. Finally, we induced interference to the WNT10B/FZD6 protein complex formation by exposure to the TGFbRI inhibitor A83-01 via inhibiting FZD6 expression, confirming its role in the WNT10B-mediated signaling activation. Conclusion Our study describes the molecular circuit of WNT10BR-mediated activation and highlight a strategy for a major improvement in T-ALL treatment.By altering FZD6-WNT10B complex formation, may provide the basis for therapeutic strategies to eradicate leukemic stem cells in patients selectively deployed depending on the underlying genetics of disease. Disclosures No relevant conflicts of interest to declare.

Download Full-text