| Autor: |
Sylvie Lalonde, Antoinette Sero, Réjane Pratelli, Guillaume Pilot, Jin Chen, Maria I Sardi, Saman A Parsa, Do-Young Kim, Biswa R Acharya, Erica V Stein, Heng-Cheng Hu, Florent Villiers, Kouji Takeda, Yingzhen Yang, Yong S Han, Rainer Schwacke, William Chiang, Naohiro Kato, Dominique Loqué, Sarah M Assmann, June M Kwak, Julian Schroeder, Seung Y Rhee, Wolf B Frommer |
| Jazyk: |
angličtina |
| Rok vydání: |
2010 |
| Předmět: |
|
| Zdroj: |
Frontiers in Physiology, Vol 1 (2010) |
| Druh dokumentu: |
article |
| ISSN: |
1664-042X |
| DOI: |
10.3389/fphys.2010.00024 |
| Popis: |
Interactions between membrane proteins and the soluble fraction are essential for signal transduction and for regulating nutrient transport. To gain insights into the membrane-based interactome, 3,852 open reading frames (ORFs) out of a target list of 8,383 representing membrane and signaling proteins from Arabidopsis thaliana were cloned into a Gateway compatible vector. The mating-based split-ubiquitin system was used to screen for potential protein-protein interactions (pPPIs) among 490 Arabidopsis ORFs. A binary robotic screen between 142 receptor-like kinases, 72 transporters, 57 soluble protein kinases and phosphatases, 40 glycosyltransferases, 95 proteins of various functions and 89 proteins with unknown function detected 387 out of 90,370 possible PPIs. A secondary screen confirmed 343 (of 387) pPPIs between 179 proteins, yielding a scale-free network (r2=0.863). Eighty of 142 transmembrane receptor-like kinases (RLK) tested positive, identifying three homomers, 63 heteromers and 80 pPPIs with other proteins. Thirty-one out of 142 RLK interactors (including RLKs) had previously been found to be phosphorylated; thus interactors may be substrates for respective RLKs. None of the pPPIs described here had been reported in the major interactome databases, including potential interactors of G protein-coupled receptors, phospholipase C, and AMT ammonium transporters. Two RLKs found as putative interactors of AMT1;1 were independently confirmed using a split luciferase assay in Arabidopsis protoplasts. These RLKs may be involved in ammonium-dependent phosphorylation of the C-terminus and regulation of ammonium uptake activity. The robotic screening method established here will enable a systematic analysis of membrane protein interactions in fungi, plants and metazoa. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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