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dc.creatorKaneko, Kentaroes_ES
dc.creatorTakamatsu, Takeshies_ES
dc.creatorInomata, Takuyaes_ES
dc.creatorOikawa, Kazusatoes_ES
dc.creatorPozueta Romero, Javieres_ES
dc.date.accessioned2020-09-22T07:53:18Z
dc.date.available2020-09-22T07:53:18Z
dc.date.issued2016
dc.identifier.issn1471-9053
dc.identifier.urihttps://hdl.handle.net/2454/38218
dc.description.abstractNucleotide pyrophosphatase/phosphodiesterases (NPPs) are widely distributed N-glycosylated enzymes that catalyze the hydrolytic breakdown of numerous nucleotides and nucleotide sugars. In many plant species, NPPs are encoded by a small multigene family, which in rice are referred to NPP1–NPP6. Although recent investigations showed that N-glycosylated NPP1 is transported from the endoplasmic reticulum (ER)–Golgi system to the chloroplast through the secretory pathway in rice cells, information on N-glycan composition and subcellular localization of other NPPs is still lacking. Computer-assisted analyses of the amino acid sequences deduced from different Oryza sativa NPP-encoding cDNAs predicted all NPPs to be secretory glycoproteins. Confocal fluorescence microscopy observation of cells expressing NPP2 and NPP6 fused with green fluorescent protein (GFP) revealed that NPP2 and NPP6 are plastidial proteins. Plastid targeting of NPP2–GFP and NPP6–GFP was prevented by brefeldin A and by the expression of ARF1(Q71L), a dominant negative mutant of ADP-ribosylation factor 1 that arrests the ER to Golgi traffic, indicating that NPP2 and NPP6 are transported from the ER–Golgi to the plastidial compartment. Confocal laser scanning microscopy and high-pressure frozen/freeze-substituted electron microscopy analyses of transgenic rice cells ectopically expressing the trans-Golgi marker sialyltransferase fused with GFP showed the occurrence of contact of Golgi-derived membrane vesicles with cargo and subsequent absorption into plastids. Sensitive and high-throughput glycoblotting/mass spectrometric analyses showed that complex-type and paucimannosidic-type glycans with fucose and xylose residues occupy approximately 80% of total glycans of NPP1, NPP2 and NPP6. The overall data strongly indicate that the trans-Golgi compartments participate in the Golgi to plastid trafficking and targeting mechanism of NPPs.en
dc.description.sponsorshipThis research was supported by the Japan Society for the Promotion of Sciences [KAKENHI Grants-in-Aid for Scientific Research (A) (15H02486) to T.M.]; the Comisión Interministerial de Ciencia y Tecnología and Fondo Europeo de Desarrollo Regional (Spain) [grants BIO2010-18239 and BIO2013-49125-C2-1-P]; the Government of Navarra [grant IIQ14067.RI1].en
dc.format.extent19 p.
dc.format.mimetypeapplication/pdfen
dc.format.mimetypeapplication/zipen
dc.language.isoengen
dc.publisherOxford University Pressen
dc.relation.ispartofPlant and Cell Physiology, 2016, 57(8), 1610–1628en
dc.rights© The Author 2016. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.comen
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/
dc.subjectGlycoproteinen
dc.subjectGolgien
dc.subjectMembrane trafficen
dc.subjectN-glycomeen
dc.subjectOryza sativaen
dc.subjectPlastid.edited-statecorrected-proofen
dc.titleN-glycomic and microscopic subcellular localization analyses of NPP1, 2 and 6 strongly indicate that trans-Golgi compartments participate in the Golgi to plastid traffic of nucleotide pyrophosphatase/phosphodiesterases in riceen
dc.typeinfo:eu-repo/semantics/articleen
dc.typeArtículo / Artikuluaes
dc.contributor.departmentIdAB - Instituto de Agrobiotecnología / Agrobioteknologiako Institutuaes
dc.rights.accessRightsinfo:eu-repo/semantics/openAccessen
dc.rights.accessRightsAcceso abierto / Sarbide irekiaes
dc.identifier.doi10.1093/pcp/pcw089
dc.relation.projectIDinfo:eu-repo/grantAgreement/ES/1PE/BIO2013-49125en
dc.relation.publisherversionhttps://doi.org/10.1093/pcp/pcw089
dc.type.versioninfo:eu-repo/semantics/publishedVersionen
dc.type.versionVersión publicada / Argitaratu den bertsioaes
dc.contributor.funderGobierno de Navarra / Nafarroako Gobernua, IIQ14067.RI1es


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© The Author 2016. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
Except where otherwise noted, this item's license is described as © The Author 2016. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com