Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network
In Arabidopsis thaliana, different types of vacuolar receptors were discovered. The AtVSR (Vacuolar Sorting Receptor) receptors are well known to be involved in the traffic to lytic vacuole (LV), while few evidences demonstrate the involvement of the receptors from AtRMR family (Receptor Membrane RI...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2016-09-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1422-0067/17/10/1661 |
| _version_ | 1811193517979992064 |
|---|---|
| author | Alessandro Occhialini Guillaume Gouzerh Gian-Pietro Di Sansebastiano Jean-Marc Neuhaus |
| author_facet | Alessandro Occhialini Guillaume Gouzerh Gian-Pietro Di Sansebastiano Jean-Marc Neuhaus |
| author_sort | Alessandro Occhialini |
| collection | DOAJ |
| description | In Arabidopsis thaliana, different types of vacuolar receptors were discovered. The AtVSR (Vacuolar Sorting Receptor) receptors are well known to be involved in the traffic to lytic vacuole (LV), while few evidences demonstrate the involvement of the receptors from AtRMR family (Receptor Membrane RING-H2) in the traffic to the protein storage vacuole (PSV). In this study we focused on the localization of two members of AtRMR family, AtRMR1 and -2, and on the possible interaction between these two receptors in the plant secretory pathway. Our experiments with agroinfiltrated Nicotiana benthamiana leaves demonstrated that AtRMR1 was localized in the endoplasmic reticulum (ER), while AtRMR2 was targeted to the trans-Golgi network (TGN) due to the presence of a cytosolic 23-amino acid sequence linker. The fusion of this linker to an equivalent position in AtRMR1 targeted this receptor to the TGN, instead of the ER. By using a Bimolecular Fluorescent Complementation (BiFC) technique and experiments of co-localization, we demonstrated that AtRMR2 can make homodimers, and can also interact with AtRMR1 forming heterodimers that locate to the TGN. Such interaction studies strongly suggest that the transmembrane domain and the few amino acids surrounding it, including the sequence linker, are essential for dimerization. These results suggest a new model of AtRMR trafficking and dimerization in the plant secretory pathway. |
| first_indexed | 2024-04-12T00:09:53Z |
| format | Article |
| id | doaj.art-e2384ed1f11846e48d4d4b4110fd5c35 |
| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-12T00:09:53Z |
| publishDate | 2016-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e2384ed1f11846e48d4d4b4110fd5c352022-12-22T03:56:00ZengMDPI AGInternational Journal of Molecular Sciences1422-00672016-09-011710166110.3390/ijms17101661ijms17101661Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi NetworkAlessandro Occhialini0Guillaume Gouzerh1Gian-Pietro Di Sansebastiano2Jean-Marc Neuhaus3Plant Biology and Crop Science, Rothamsted Research, Harpenden, AL5 2JQ Herts, UKLaboratory of Cell and Molecular Biology, Institute of Biology, University of Neuchâtel, Rue Emile-Argand 11, CH-2009 Neuchâtel, SwitzerlandDISTEBA, Department of Biological and Environmental Sciences and Technologies, University of Salento, Campus Ecotekne, 73100 Lecce, ItalyLaboratory of Cell and Molecular Biology, Institute of Biology, University of Neuchâtel, Rue Emile-Argand 11, CH-2009 Neuchâtel, SwitzerlandIn Arabidopsis thaliana, different types of vacuolar receptors were discovered. The AtVSR (Vacuolar Sorting Receptor) receptors are well known to be involved in the traffic to lytic vacuole (LV), while few evidences demonstrate the involvement of the receptors from AtRMR family (Receptor Membrane RING-H2) in the traffic to the protein storage vacuole (PSV). In this study we focused on the localization of two members of AtRMR family, AtRMR1 and -2, and on the possible interaction between these two receptors in the plant secretory pathway. Our experiments with agroinfiltrated Nicotiana benthamiana leaves demonstrated that AtRMR1 was localized in the endoplasmic reticulum (ER), while AtRMR2 was targeted to the trans-Golgi network (TGN) due to the presence of a cytosolic 23-amino acid sequence linker. The fusion of this linker to an equivalent position in AtRMR1 targeted this receptor to the TGN, instead of the ER. By using a Bimolecular Fluorescent Complementation (BiFC) technique and experiments of co-localization, we demonstrated that AtRMR2 can make homodimers, and can also interact with AtRMR1 forming heterodimers that locate to the TGN. Such interaction studies strongly suggest that the transmembrane domain and the few amino acids surrounding it, including the sequence linker, are essential for dimerization. These results suggest a new model of AtRMR trafficking and dimerization in the plant secretory pathway.http://www.mdpi.com/1422-0067/17/10/1661AtRMRPA domainRING-H2Ser-Rich domainlinkertransmembraneplant secretory pathwaytrans-Golgi networkendoplasmic reticulumdimerizationlaser scanning confocal microscopyBimolecular Fluorescent ComplementationArabidopsis thalianaNicotiana benthamiana |
| spellingShingle | Alessandro Occhialini Guillaume Gouzerh Gian-Pietro Di Sansebastiano Jean-Marc Neuhaus Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network International Journal of Molecular Sciences AtRMR PA domain RING-H2 Ser-Rich domain linker transmembrane plant secretory pathway trans-Golgi network endoplasmic reticulum dimerization laser scanning confocal microscopy Bimolecular Fluorescent Complementation Arabidopsis thaliana Nicotiana benthamiana |
| title | Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network |
| title_full | Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network |
| title_fullStr | Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network |
| title_full_unstemmed | Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network |
| title_short | Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network |
| title_sort | dimerization of the vacuolar receptors atrmr1 and 2 from arabidopsis thaliana contributes to their localization in the trans golgi network |
| topic | AtRMR PA domain RING-H2 Ser-Rich domain linker transmembrane plant secretory pathway trans-Golgi network endoplasmic reticulum dimerization laser scanning confocal microscopy Bimolecular Fluorescent Complementation Arabidopsis thaliana Nicotiana benthamiana |
| url | http://www.mdpi.com/1422-0067/17/10/1661 |
| work_keys_str_mv | AT alessandroocchialini dimerizationofthevacuolarreceptorsatrmr1and2fromarabidopsisthalianacontributestotheirlocalizationinthetransgolginetwork AT guillaumegouzerh dimerizationofthevacuolarreceptorsatrmr1and2fromarabidopsisthalianacontributestotheirlocalizationinthetransgolginetwork AT gianpietrodisansebastiano dimerizationofthevacuolarreceptorsatrmr1and2fromarabidopsisthalianacontributestotheirlocalizationinthetransgolginetwork AT jeanmarcneuhaus dimerizationofthevacuolarreceptorsatrmr1and2fromarabidopsisthalianacontributestotheirlocalizationinthetransgolginetwork |