Cargando…

The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress

Alternative splicing (AS) is an important molecular mechanism by which single genes can generate multiple mRNA isoforms. We reported previously that, in Oryza sativa, the cyclophilin 19-4 (OsCYP19-4.1) transcript was significantly upregulated in response to cold stress, and that transgenic plants we...

Descripción completa

Detalles Bibliográficos
Autores principales: Lee, Areum, Lee, Sang Sook, Jung, Won Yong, Park, Hyun Ji, Lim, Bo Ra, Kim, Hyun-Soon, Ahn, Jun Cheul, Cho, Hye Sun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4964526/
https://www.ncbi.nlm.nih.gov/pubmed/27447607
http://dx.doi.org/10.3390/ijms17071154
_version_ 1782445130393845760
author Lee, Areum
Lee, Sang Sook
Jung, Won Yong
Park, Hyun Ji
Lim, Bo Ra
Kim, Hyun-Soon
Ahn, Jun Cheul
Cho, Hye Sun
author_facet Lee, Areum
Lee, Sang Sook
Jung, Won Yong
Park, Hyun Ji
Lim, Bo Ra
Kim, Hyun-Soon
Ahn, Jun Cheul
Cho, Hye Sun
author_sort Lee, Areum
collection PubMed
description Alternative splicing (AS) is an important molecular mechanism by which single genes can generate multiple mRNA isoforms. We reported previously that, in Oryza sativa, the cyclophilin 19-4 (OsCYP19-4.1) transcript was significantly upregulated in response to cold stress, and that transgenic plants were cold tolerant. Here we show that, under cold stress, OsCYP19-4 produces eight transcript variants by intron retention and exon skipping, resulting in production of four distinct protein isoforms. The OsCYP19-4 AS isoforms exhibited different cellular localizations in the epidermal cells: in contrast to OsCYP19-4.1, the OsCYP19-4.2 and OsCYP19-4.3 proteins were primarily targeted to guard and subsidiary cells, whereas OsCYP19-4.5, which consists largely of an endoplasmic reticulum (ER) targeting signal, was co-localized with the RFP-BiP marker in the ER. In OsCYP19-4.2, the key residues of the PPIase domain are altered; consistent with this, recombinant OsCYP19-4.2 had significantly lower PPIase activity than OsCYP19-4.1 in vitro. Specific protein-protein interactions between OsCYP19-4.2/3 and AtRCN1 were verified in yeast two-hybrid (Y2H) and bimolecular fluoresence complementation (BiFC assays), although the OsCYP19-4 isoforms could not bind each other. Based on these results, we propose that two OsCYP19-4 AS isoforms, OsCYP19-4.2 and OsCYP19-4.3, play roles linking auxin transport and cold stress via interactions with RCN1.
format Online
Article
Text
id pubmed-4964526
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-49645262016-08-03 The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress Lee, Areum Lee, Sang Sook Jung, Won Yong Park, Hyun Ji Lim, Bo Ra Kim, Hyun-Soon Ahn, Jun Cheul Cho, Hye Sun Int J Mol Sci Article Alternative splicing (AS) is an important molecular mechanism by which single genes can generate multiple mRNA isoforms. We reported previously that, in Oryza sativa, the cyclophilin 19-4 (OsCYP19-4.1) transcript was significantly upregulated in response to cold stress, and that transgenic plants were cold tolerant. Here we show that, under cold stress, OsCYP19-4 produces eight transcript variants by intron retention and exon skipping, resulting in production of four distinct protein isoforms. The OsCYP19-4 AS isoforms exhibited different cellular localizations in the epidermal cells: in contrast to OsCYP19-4.1, the OsCYP19-4.2 and OsCYP19-4.3 proteins were primarily targeted to guard and subsidiary cells, whereas OsCYP19-4.5, which consists largely of an endoplasmic reticulum (ER) targeting signal, was co-localized with the RFP-BiP marker in the ER. In OsCYP19-4.2, the key residues of the PPIase domain are altered; consistent with this, recombinant OsCYP19-4.2 had significantly lower PPIase activity than OsCYP19-4.1 in vitro. Specific protein-protein interactions between OsCYP19-4.2/3 and AtRCN1 were verified in yeast two-hybrid (Y2H) and bimolecular fluoresence complementation (BiFC assays), although the OsCYP19-4 isoforms could not bind each other. Based on these results, we propose that two OsCYP19-4 AS isoforms, OsCYP19-4.2 and OsCYP19-4.3, play roles linking auxin transport and cold stress via interactions with RCN1. MDPI 2016-07-19 /pmc/articles/PMC4964526/ /pubmed/27447607 http://dx.doi.org/10.3390/ijms17071154 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lee, Areum
Lee, Sang Sook
Jung, Won Yong
Park, Hyun Ji
Lim, Bo Ra
Kim, Hyun-Soon
Ahn, Jun Cheul
Cho, Hye Sun
The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress
title The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress
title_full The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress
title_fullStr The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress
title_full_unstemmed The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress
title_short The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress
title_sort oscyp19-4 gene is expressed as multiple alternatively spliced transcripts encoding isoforms with distinct cellular localizations and ppiase activities under cold stress
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4964526/
https://www.ncbi.nlm.nih.gov/pubmed/27447607
http://dx.doi.org/10.3390/ijms17071154
work_keys_str_mv AT leeareum theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT leesangsook theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT jungwonyong theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT parkhyunji theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT limbora theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT kimhyunsoon theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT ahnjuncheul theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT chohyesun theoscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT leeareum oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT leesangsook oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT jungwonyong oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT parkhyunji oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT limbora oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT kimhyunsoon oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT ahnjuncheul oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress
AT chohyesun oscyp194geneisexpressedasmultiplealternativelysplicedtranscriptsencodingisoformswithdistinctcellularlocalizationsandppiaseactivitiesundercoldstress