Document type
Journal articles
Document subtype
Full paper
Title
Functional characterization reveals the importance of Arabidopsis ECA4 and EPSIN3 in clathrin mediated endocytosis and wall structure in apical growing cells
Participants in the publication
Rita Teresa Teixeira (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Dep. Biologia Vegetal
BioISI
Dario Marchese (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Dep. Biologia Vegetal
BioISI
Patrick J. Duckney (Author)
UNIVERSITY OF DURHAM
Fernando Vaz Dias (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Ana P Carapeto (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Mariana Louro (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Marta Sousa Silva (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Carlos Cordeiro (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Mário S. Rodrigues (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Rui Malhó (Author)
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Summary
Localized clathrin mediated endocytosis is vital for secretion and wall deposition in apical growing plant cells. Adaptor and signalling proteins, along with phosphoinositides, are known to play a regulatory, yet poorly defined role in this process. Here we investigated the function of Arabidopsis ECA4 and EPSIN3, putative mediators of the process, in pollen tubes and root hairs.\nHomozygous eca4 and epsin3 plants exhibited altered pollen tube morphology (in vitro) and self-pollination led to fewer seeds and shorter siliques. These effects were augmented in eca4/epsin3 double mutant and quantitative polymerase chain reaction data revealed changes in phosphoinositide metabolism and flowering genes suggestive of a synergistic action. No visible changes were observed in root morphology, but atomic force microscopy in mutant root hairs showed altered structural stiffness.\nImaging and FRET-FLIM analysis of ECA4 and EPSIN3 X-FP constructs revealed that both proteins interact at the plasma membrane but exhibit slightly different intracellular localization. FT-ICR-MS metabolomic analysis of mutant cells showed changes in lipids, amino acids and carbohydrate composition consistent with a role in secretion and growth.\nCharacterization of double mutants of eca4 and epsin3 with phospholipase C genes (plc5, plc7) indicates that phosphoinositides (e.g. PtdIns(4,5)P2) are fundamental for a combined and complementary role of ECA4-EPSIN3 in cell secretion.
Date of Submisson/Request
2024-08-13
Date of Acceptance
2024-10-26
Date of Publication
2024-11-18
Institution
FACULDADE DE CIÊNCIAS DA UNIVERSIDADE DE LISBOA
Where published
New Phytologist
Publication Identifiers