Os03g0356414
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Contents
Annotated Information
Function
OsDIS1 Negatively Regulates the Drought Response in Rice To explore the biological function of OsDIS1, we created OsDIS1 overexpression and RNAi constructs, which were transformed into the japonica cv Nipponbare by Agrobacterium tumefaciens-mediated transformation (Qu et al., 2006). The expression of OsDIS1 in the overexpression homozygous lines was first analyzed by RNA gel blot using the whole coding sequence of OsDIS1 as the probe . Four independent homozygous transgenic lines (OX 9-4-2, OX 17-2-8, OX 35-3-2, and OX 44-1-1) with obvious high OsDIS1 expression were chosen for further western blot analysis. In all four lines, a strong accumulation of the TAP-OsDIS1 fusion protein was detected by peroxidase anti-peroxidase antibody . The expression of OsDIS1 in the OsDIS1 RNAi homozygous lines was first analyzed by reverse transcription (RT)-PCR and then confirmed by real-time PCR. Four lines (RNAi 2-1-2, RNAi 7-1-1, RNAi 9-1-3, and RNAi 22-1-6) with significantly decreased expression of OsDIS1 were chosen for further analysis. Real-time PCR analysis revealed that the expression of OsDIS1’s five paralogs was not affected in both overexpression and RNAi lines using five pairs of gene-specific primers. The drought response of the OsDIS1 overexpression and RNAi lines was evaluated with 4-week-old transgenic and wild-type Nipponbare plants. Under our greenhouse conditions and without drought stress, the growth of both types of OsDIS1 transgenic plants was similar to that of the wild-type plants. Then, these plants were removed from the trays and kept on a bench to induce drought stress.After 6 d without watering, the leaves of the OsDIS1 overexpression plants began to roll but the leaves of the OsDIS1 RNAi plants and wild-type plants had no obvious changes . After 7 d without watering, the leaves of all overexpression plants were curled, while those of the wild-type plants were only beginning to show the drought-stress phenotypes . On day 9, all of the overexpression plants and most of the wild-type plants had wilted, while the leaves of the RNAi plants had just begun to curl. At the end of 9 d, all plants were moved back to trays with water. After 4 d of recovery in water, about 75% to 90% of the RNAi plants recovered from the stress and started to grow , but only about 50% of the wild-type plants and 20% to 30% of the overexpression plants survived.These results demonstrate that OsDIS1 has a negative role in drought tolerance in rice.
Expression
To confirm the microarray expression results for OsDIS1, real-time PCR analysis was conducted with the OsDIS1 gene-specific primers using RNA isolated from 7-d-old seedlings before and after drought treatment. In the reactions, OsDREB2Awas used as a positive control for the drought treatment. Similar to OsDREB2A, OsDIS1 was highly induced 24 h after the drought treatment (Fig. 1B), suggesting that OsDIS1 is involved in the drought response in rice. OsDIS1 Is Localized Predominantly in the Nucleus To investigate the subcellular localization of OsDIS1,the full-length cDNA of OsDIS1 was introduced into both pGDG and pGDR vectors, which contain a cauliflower mosaic virus promoter-driven GFP and the DsRed gene, respectively. Subcellular localization assays were conducted in both N.benthamiana leaves and rice protoplasts. Localization of the fusion protein was then determined by visualization with a confocal microscope, and the position of the nucleus of the N. benthamiana epidermal cell was confirmed with 4',6-diamidino-2-phenylindole (DAPI) staining.Transient expression assays in N. benthamiana leaves indicated that the GFP-OsDIS1 fusion protein was localized predominantly in the nucleus, as well as some in the cytoplasm and plasma membrane , which is consistent with the distribution of SINAT5 in Arabidopsis.The localization of OsDIS1 in N. benthamiana leaves was confirmed by its subcellular localization in rice protoplasts.
Evolution
Among the analyzed 378 RING finger-type and 76 U-box-type E3 ubiquitin ligase genes in rice, a drought induced C3HC4 RING finger gene (LOC_Os03g24040) was identified.The deduced protein of this gene contains conserved RING finger and zinc finger motifs and has high sequence identity (82%) with SINAT5 in Arabidopsis,which 2002), which was identified as a homolog of Drosophila SINA.A genome-wide search identified five paralogous SINA proteins of OsDIS1 in the rice genome. The amino acid identity between OsDIS1 and its paralogs is 91% with Os07g46560, 83% with Os02g19140, 83% with Os05g14860, 83% with Os01g13370, and 71% with Os02g03620. Further alignment between rice and Arabidopsis SINA proteins revealed that except for the N terminus, all the SINA members in rice and Arabidopsis had highly conserved sequences, and all contained the conserved RING finger and zinc finger motifs.Phylogenetic analysis showed that each of the following protein pairs belongs to the same clade: SINAT1 and SINAT2, SINAT4 and SINAT5, and OsDIS1 and Os07g46560; this supports the inference of a possible duplication of these genes in both genomes. Based on pairwise distance analysis, OsDIS1 has a minimal distance of 0.0725 with Os0746560 in rice and 0.2624 with SINAT5 in Arabidopsis,suggesting that Os07g46560 is the closest paralog of OsDIS1 in rice and SINAT5 is the closest ortholog of OsDIS1 in Arabidopsis.
Labs working on this gene
State Laboratory for Biology of Plant Diseases and Insect Pests; Institute of Plant Protection; Chinese Academy of Agricultural Sciences; Beijing, China State Key Laboratory of Plant Genomics; National Center for Plant Gene Research; Institute of Genetics and Developmental Biology; Chinese Academy of Sciences; Beijing, China Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization; Hunan Agricultural University; Changsha Hunan, China; Department of Plant Pathology;Ohio State University; Columbus, OH USA;Department of Applied Plant Sciences Technology, Kangwon National University；Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University,Seoul 120–749，Korea.
References
1. Ning,Yuese and Jantasuriyarat.The SINA E3 ligase OsDIS1 negatively regulates drought response in rice.Plant physiology.2011,157:242--255； 2. Ning, Yuese and Xie.OsDIS1-mediated stress response pathway in rice.Plant Signal Behav.2011,6:1684--1686； 3. Yamada, Yasuyuki and Koyama.Basic helix-loop-helix transcription factors and regulation of alkaloid biosynthesis.Plant Signal Behav.2011,6:1627--1630； 4. Deikman, Jill and Petracek.Drought tolerance through biotechnology: improving translation from the laboratory to farmers’ fields.Current opinion in biotechnology.2012,23:243--250； 5. Lim, Sung Don and Hwang.Comprehensive Analysis of the Rice RING E3 Ligase Family Reveals Their Functional Diversity in Response to Abiotic stress.DNA research.2013,20:299--314； 6. Cho, Seok Keun and Ryu.The Arabidopsis RING E3 ubiquitin ligase AtAIRP2 plays combinatory roles with AtAIRP1 in abscisic acid-mediated drought stress responses.Plant physiology.2011,157:2240--2257
Structured Information
| Gene Name |
Os03g0356414 |
|---|---|
| Description |
Similar to Ubiquitin ligase SINAT5 (EC 6.3.2.-) (Seven in absentia homolog 5). Splice isoform 2 |
| Version |
NM_001186491.1 GI:297722112 GeneID:9266295 |
| Length |
3760 bp |
| Definition |
Oryza sativa Japonica Group Os03g0356414, complete gene. |
| Source |
Oryza sativa Japonica Group ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
| Chromosome | |
| Location |
Chromosome 3:14184730..14188489 |
| Sequence Coding Region |
14185089..14185409,14185874..14186260,14186613..14186810 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008396:14184730..14188489 source=RiceChromosome03 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008396:14184730..14188489 source=RiceChromosome03 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atggcctcagttacttatattgatgactccggttctgaggtaattgatcctccaaagactgaggtgctggatgttaccgaacttgctggtgatcctgttccgcattcaccaaaaccaaatgtggtagtttctagcagtgtgcgtgaactgcttgaatgtccagtctgcctgagcgcaatgtatcctcccattcatcagtgctctaatggtcatacattgtgctctggatgcaaaccaagggttcacaatcgctgtccaacttgtaggcatgaactgggtaatattagatgtcttgctctcgagaaggtggctgcgtcgcttgaacttccatgcaagtaccagaacttcgggtgtgtaggcatttatccttactactgcaagctgaagcatgagtcacagtgccaatataggccttatagctgtccatatgctggatctgaatgcacagttgctggtgacattccatacttggtgaatcacttgaaagatgaccacaaggttgacatgcataatggctgcaccttcaaccatcgctatgtcaagtcgaatcctcatgaagttgagaatgccacctggatgcttacggttttcagctgctttggccagtacttctgcctacatttcgaagcatttcagctggggatggcacctgtgtacatcgccttcctgaggttcatgggtgatgacttggaagcaaagaactacagctacagcctggaggtaggaggcactggccgcaaaatgatctggcaaggggttccccggagcatcagagacagccatcggaaggtccgggatagctatgatgggcttatcatccaacggaacatggccttgttcttctctggtggagaaaggaaggagctcaaattgcgggtcactgggagaatttggaaggaacagtga</cdnaseq> |
| Protein Sequence |
<aaseq>MASVTYIDDSGSEVIDPPKTEVLDVTELAGDPVPHSPKPNVVVS SSVRELLECPVCLSAMYPPIHQCSNGHTLCSGCKPRVHNRCPTCRHELGNIRCLALEK VAASLELPCKYQNFGCVGIYPYYCKLKHESQCQYRPYSCPYAGSECTVAGDIPYLVNH LKDDHKVDMHNGCTFNHRYVKSNPHEVENATWMLTVFSCFGQYFCLHFEAFQLGMAPV YIAFLRFMGDDLEAKNYSYSLEVGGTGRKMIWQGVPRSIRDSHRKVRDSYDGLIIQRN MALFFSGGERKELKLRVTGRIWKEQ</aaseq> |
| Gene Sequence |
<dnaseqindica>3081..3401#2230..2616#1680..1877#aaaaaacgcgagaagagagagagagaagcttttaattcgaattctctctctcatcacggcttcttctgcttctccttctcctcccccctcatccctcctccgccggtagccgccggccgggagcgagcagcgagcgagaccggagcccgccggagccggtctcgctcccccctcctcccttgttcgcgccacccccgaggtacatcccgccgatctctctctttctctctctcgttggtctgctctgtgattttttttttttttgccaaggctgttctttatttggcacagagattgttttttttttccttctttgcgggtgtccggttcgagtccacggggtttgactctgccggaaatggttcgaatgcggcggccggagggcggaggaggggttttggggggcgttcttggtgggttggtttgcgccgagtctttgttaatttgttagggttcttgatctgatgcggcttctcgttgccgggatgcgcttgcctcatggctgaattcaaacaggccggtctctctctccctccctctctcccgtcagccgtgtgctttgaccgagtaagtttttgaacttcgattggttgagctgggggttcttgactttttcttactccacgaatgttcctttttgcacaatggggctactaatgaatggaaagctagagatggtgtgcctaatggtgatgtgccgtcattgtgttagctcggatttgtactgcgaatttgcgagctcacatgctcagctgatgatgtgttgatccagggagtatatataattggttggtagagtacttgctagattgtgctcacgggtctcatttcaaagttcagtatgaaatcttggttgtaaagtagtactagtgttttgttaatcgttagggagtacttctcctgcatttttcttaattttttccctttaaaaactgtaggtggatttgttattagtaaagctgatttgtgccaaatagaaattctgttttatccgcagtaaggtactgtaaacagtacttgcaggcactagggtttattggcaaggttttctatgttacattttaataaatgccacctttttcttcgtcaaaagaagccacggttataaagaaaaagtgaagcattaattgatgtaaatgcacgaactacaaatttccacaaatataaaccttttaataaaatccacaaatgaaagaaaaatgcagttcacacaccaatctctttctcaaaatttacacttttcttataacaggtatgctaaattttcttaatacaaagcatatacaaaacatcacaagcactatagacattcatgacatatctgttgcataggacaatatgcagtatgaagacctggctggtcaaaacttccgatgcacatgtgaggccctatccatgtgctttcttgatttctgctcctttatggccctatctccttcaaagcccttaaacacttcaacagtgtccaatcatactatgagactgtatggatatgggcatatggctatacttttgatgaggcaatgattttttttgttaataatagtgttaataggaagtgttgaattggttttttaaaaggaagtatttaagcattgaatttgtcatttattttgactcactaataacacagatagttctctttcctagcaggaccccttacaatatctagtgtatttatggcctcagttacttatattgatgactccggttctgaggtaattgatcctccaaagactgaggtgctggatgttaccgaacttgctggtgatcctgttccgcattcaccaaaaccaaatgtggtagtttctagcagtgtgcgtgaactgcttgaatgtccagtctgcctgagcgcaatgtatcctcccattcatcaggtgcacatataccacttattgaatcttttttgcacttaaaaaggatattttcttgtatggaagtcacttgggaaacaaaatgcgtgtttcatttaattgcattgcttttactatcttgaatcctgctatcatatttgtagtctggcctgatctatttgaacttatctagatgatttcctggtttaaaacggtcgttctgagctgctttttcacatatgttctgttccactaccaaattgttacccatgtatttaagcaccaaaaaagaagttagctaagatagtatgtacaatatgcacattgttgtactatgttgtaagttgtaaccccaactctttttctgctaatgcagtgctctaatggtcatacattgtgctctggatgcaaaccaagggttcacaatcgctgtccaacttgtaggcatgaactgggtaatattagatgtcttgctctcgagaaggtggctgcgtcgcttgaacttccatgcaagtaccagaacttcgggtgtgtaggcatttatccttactactgcaagctgaagcatgagtcacagtgccaatataggccttatagctgtccatatgctggatctgaatgcacagttgctggtgacattccatacttggtgaatcacttgaaagatgaccacaaggttgacatgcataatggctgcaccttcaaccatcgctatgtcaagtcgaatcctcatgaagttgagaatgccacctggatgcttacggtatcccctcgttaactccaccatgttagttattgtgaagtatatcactgatgtttacttatgcaaagattggttcacttgtctgctaagatagtccatatttctgtactgagttttatgagcctttctgtaaaaataaacattggttcacttgtacagtatgattgcactctgtacttttcatgtcaactgaggatgctgatcatgcagcatagttattggtctaaaataatctctcacctatgaagaaactatgcatcagaatatccatctgcttccatttccaaaaaaaaaaaaaaaaactttgtctgctttacagcacaataatagcatttggacagggctcttgatctgcaaccttttcttgctgctttctaagtgcaatataaaattaagaatttccttctaatctgttcttttccattgaacaaaatggactaagtaactataatgcttccatctaggttttcagctgctttggccagtacttctgcctacatttcgaagcatttcagctggggatggcacctgtgtacatcgccttcctgaggttcatgggtgatgacttggaagcaaagaactacagctacagcctggaggtaggaggcactggccgcaaaatgatctggcaaggggttccccggagcatcagagacagccatcggaaggtccgggatagctatgatgggcttatcatccaacggaacatggccttgttcttctctggtggagaaaggaaggagctcaaattgcgggtcactgggagaatttggaaggaacagtgaaaataaaatgtgaaatgatctgttcatcgttcttaacccttgcatgaacctatgtatcatcacagtgcgagaattatagccattcataggcagtgactctaaaatgaagacttgtaatgttattagcttgttttagctaccatcttctgttagattggtgcctgtggatgactagatgacagtcatgtagaccagagtggcactattgtagaattcctggcaaacttctctttgccttgaactgcttgttgagttgccattctgtggtatagggaaatctaatggcaattcatgagatgaatgtgttgaactctttttcatgtttccagcatatttctagcttctttgttcattctttc</dnaseqindica> |
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