PrMATE1 Is Differentially Expressed in Radiata Pine Exposed to Inclination and the Deduced Protein Displays High Affinity to Proanthocyanidin Substrates by a Computational Approach
The response to inclination in plants is an attractive and extensively studied biological process. The most commonly held theory proposes a differential growth in stem tissue due to unequal auxin redistribution. Further evidence proposed that flavonoids act as molecular regulators of auxin distribution or flux. It is well known that flavonoids affect auxin distribution, but how intracellular concentration is controlled during the gravitropic response in woody species is still unknown. The MATE family has been widely studied, however the molecular basis of flavonoids transport is still poorly understood. Here, we identified and characterized a full-length cDNA from radiate pine encoding a putative MATE protein. Transcript abundance analysis showed that PrMATE1 is expressed in a spatial and temporal manner in inclined stems. Additionally, PrMATE1 fused to GFP is mainly localized in the vacuolar membrane. A 3D protein model showed 12 transmembrane helices and an open cavity. The protein–ligand interaction was evaluated; favourable binding affinity energies were obtained and suggested epicatechin 3′-O-glucoside as the best putative ligand. In silico mutagenesis analysis was used to identify five residues as important to protein–ligand interaction. The data provide a dynamic view of interaction between PrMATE1 and their putative ligands at the molecular scale.
KeywordsMATE transporter Expression profile Molecular modelling Molecular dynamics simulations Pinus radiata Inclination stress
This work has been supported by PAI/Academia Nº 79140027, FONDECYT N° 11121170, and FONDECYT N° 1150964. PR acknowledges ‘Núcleo Científico Multidisciplinario’ from Universidad de Talca. Authors would like to thank the reviewers for their highly valuable comments.
Compliance with Ethical Standards
Conflict of interest
The authors declares that they have no conflict of interest.
- Debeaujon I, Peeters AJ, Leon-Kloosterziel KM, Koornneef M (2001) The TRANSPARENT TESTA12 gene of Arabidopsis encodes a multidrug secondary transporter-like protein required for flavonoid sequestration in vacuoles of the seed coat endothelium. Plant Cell 13:853–871CrossRefPubMedPubMedCentralGoogle Scholar
- Espinoza A, Contreras R, Zúñiga GE, Herrera R, Moya-León MA, Norambuena L, Handford M (2016) FcLDP1, a gene encoding a late embryogenesis abundant (LEA) domain protein, responds to brassinosteroids and abscisic acid during the development of fruits in Fragaria chiloensis. Front Plant Sci 7:788CrossRefPubMedPubMedCentralGoogle Scholar
- Friesner RA, Banks JL, Murphy RB, Halgren TA, Klicic JJ, Mainz DT, Repasky MP, Knoll EH, Shelley M, Perry JK, Shaw DE, Francis P, Shenkin PS (2004) Glide: A New Approach for Rapid, Accurate Docking and Scoring. 1. Method and Assessment of Docking Accuracy. J Med Chem 47:1739–1749CrossRefGoogle Scholar
- Gomez R, Gonzalez J, Herrera R, Ramos P (2017) MYB Transcription Factors and a Putative Flavonoid Transporter ABCC-Like are Differentially Expressed in Radiata Pine Seedlings Exposed to Inclination. J Plant Growth Regul 1–12Google Scholar
- Magalhaes JV, Liu J, Guimarães CT, Lana UG, Alves VM, Wang YH, Schaffert RE, Hoekenga OA, Piñeros MA, Shaff JE, Klein PE, Carneiro NP, Coelho CM, Trick HN, Kochian LV (2007) A gene in the multidrug and toxic compound extrusion (MATE) family confers aluminum tolerance in sorghum. Nat Genet 39:1156–1161CrossRefPubMedGoogle Scholar
- Marinova K, Pourcel L, Weder B, Schwarz M, Barron D, Routaboul JM, Debeaujon I, Klein M (2007) The Arabidopsis MATE transporter TT12 acts as a vacuolar flavonoid/H+-antiporter active in proanthocyanidin-accumulating cells of the seed coat. Plant Cell 19:2023–2038CrossRefPubMedPubMedCentralGoogle Scholar
- Mathews H, Clendennen SK, Caldwell CG, Liu XL, Connors K, Matheis N, Schuster DK, Menasco DJ, Wagoner W, Lightener J, Wagner DR (2003) Activation tagging in tomato identifies a transcriptional regulator of anthocyanin biosynthesis, modification, and transport. Plant Cell 15:1689–1703CrossRefPubMedPubMedCentralGoogle Scholar
- Pineau C, Loubet S, Lefoulon C, Chalies C, Fizames C, Lacombe B, Ferrand M, Loudet O, Berthomieu P, Richard O (2012) Natural variation at the FRD3 MATE transporter locus reveals cross-talk between Fe homeostasis and Zn tolerance in Arabidopsis thaliana. PLoS Genet 8:e1003120CrossRefPubMedPubMedCentralGoogle Scholar
- Schrödinger (2015) Schrödinger Release 2015-1: LigPrep. Schrödinger, LLC, New YorkGoogle Scholar
- Shitan N, Minami S, Morita M, Hayashida M, Ito S, Takanashi K, Omote H, Moriyama Y, Sugiyama A, Goossens A, Moriyasu M, Yazaki K (2014) Involvement of the leaf-specific multidrug and toxic compound extrusion (MATE) transporter Nt-JAT2 in vacuolar sequestration of nicotine in Nicotiana tabacum. PLoS ONE 9:e108789CrossRefPubMedPubMedCentralGoogle Scholar