Os10g0445400

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Annotated Information

OsHCI1 encodes a RING finger E3 ligases, which is specifically induced by heat and cold stress. OsHCI1 can drive nuclear export of multiple protein substrate, and the heterologous overexpression of Arabidopsis can enhance acquired-thermotolerance.

Function

OsHCI1 (Oryza sativa Heat and Cold Induced 1) encodes a 246-amino acid protein with a predicted molecular mass of 28.8kDa and harbours a single RING-HC domain in its C-terminal region. It is a rice RING domain E3 ligase, and is highly induced under heat and cold stress conditions, which can lead to adverse outcomes in plant cell functions, including alterations in cellular composition of membrane fluidity and permeability, enzyme activity, metabolism, production of active oxygen species, and gene expression.

OsHCI1 dynamically moves from the cytoplasm to the nucleus along cytoskeletal tracts under heat shock conditions. OsHCI1 interacts with six substrate proteins and mediates subcellular trafficking of nuclear proteins to the cytoplasm via monoubiquitination.

Arabidopsis overexpressing OsHCI1-EYFP exhibits a heat-tolerant phenotype, suggesting an important role of this protein in the regulation of heat-generated signals in plants.

Expression

The finding that OsHCI1 gene expression patterns are specifically and somewhat rapidly increased by heat and cold stresses but not by salt and drought stresses indicates that the gene is associated closely with thermal stress in rice. And OsHCI1 rapidly responds to hormone treatments,too. (Fig.1)

Fig.1. Expression levels of OsHCl1 in rice plants subjected to four abiotic stresses and four hormonal treatments.

A Y2H screen was performed to identify 6 proteins that interact with OsHCI1. To confirm these positive interactions with OsHCI1, full-length coding sequences of the top six genes, which exhibited strong α-galactosidase activity, were cloned into GAL4 activation domain, respectively. Full-length OsHCI1 and each interacting protein were co-transformed into the Y2H Gold strain and grown on QDO/X/A medium(Fig.2).

Fig2.Identification of OsHCI1 interaction with six proteins.

This study also examined the expression patterns of the interacting partner genes with OsHCI1 under two different heat stresses via semi-quantitative RT-PCR with rice seedlings treated by basal or acquired heat shock treatments (Fig.3). These results suggest that heat shock results in high expression of the OsHCI1 transcript or protein, which can affect the transcript levels of its interacting genes.

Fig3. Expression patterns of the response of interacting protein genes with OsHCI1 under heat treatment.

Localization

BiFC technology was employed to visualize the interactions between OsHCI1 and each of the interaction partners in living cells . Full-length coding sequences of OsHCI1 and each of the six interacting protein genes were cloned into the 35S-HA-SPYCE(M) and 35S-c-myc-SPYNE(R)173 vectors, respectively. All of the YFP signals except that of OsPSA7 appeared to associate with the cytoplasm and nucleus; however, the OsPGLU1-, OsbHLH065-, and OsGRP1-DsRed2 alone protein signals were detected only in the nucleus (Fig. 4B–D). In contrast, the OsHCI1 BiFC complex with OsPSA7 was localized to the cytoplasm with a punctuate complex (Fig. 5).

Fig. 4. Subcellular localization of six interacting proteins.

Fig.5. BiFC assay for six substrate proteins confirms the interaction with OsHCI1 in living cells.

Labs working on this gene

1.Department of Applied Plant Sciences, Kangwon National University, Chuncheon 200-713, Korea.

Structured Information

Gene Name	Os10g0445400
Description	Zinc finger, RING-type domain containing protein
Version	NM_001071244.2 GI:297610563 GeneID:4348743
Length	2212 bp
Definition	Oryza sativa Japonica Group Os10g0445400, 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	Chromosome 10
Location	Chromosome 10:16541690..16543901
Sequence Coding Region	16541849..16542062,16542867..16543089,16543187..16543253,16543374..16543610
Expression	GEO Profiles:Os10g0445400
Genome Context	<gbrowseImage1> name=NC_008403:16541690..16543901 source=RiceChromosome10 preset=GeneLocation </gbrowseImage1>
Gene Structure	<gbrowseImage2> name=NC_008403:16541690..16543901 source=RiceChromosome10 preset=GeneLocation </gbrowseImage2>
Coding Sequence	<cdnaseq>atgagctgtgagttcttcctcccttcgccggtcgcggctcgtggacggtggtggtggtggggggtttgcgtgtggttcatctctctcttttgcagggcgtcggagttcttgcgggattacgacggggcggtgatccagatgcggatggcgtacagcgccgtcgcgcacttcctcgtgcagtggatcgactgcaagctcgccggcgcgctcggcctcctcaagatcatgatctacaaggtgtacgccgatggcaccacggctctgccggagtgggagagggaggccagcatcaggcaattctacggtgtcatcttcccgtcgctgctccagctgccgagtgggataactgaattggacgacaggaagcagaggaggctgtgccttcagaagttcaggaaggtggaggagagggtctcggaggtggatttggagagggagctcgagtgcggcatctgcctcgaggtgaatgccaagattgtgctgcccgattgcgcgcactcgctgtgcatgagatgcttcgaggattggaacaccaaatcaaagtcgtgccccttctgccgcgcctgcctcaagaaggtgaatccgagcagcctgtggttgtacaccgacgaccgcgatgttgtggatatggatacgttgactagggagaacattaggcgcctgttcatgttcataagtaagcttccacttgtagtgctccatgtggttgaccttgacatttacgagtaccgtatcaagtga</cdnaseq>
Protein Sequence	<aaseq>MSCEFFLPSPVAARGRWWWWGVCVWFISLFCRASEFLRDYDGAV IQMRMAYSAVAHFLVQWIDCKLAGALGLLKIMIYKVYADGTTALPEWEREASIRQFYG VIFPSLLQLPSGITELDDRKQRRLCLQKFRKVEERVSEVDLERELECGICLEVNAKIV LPDCAHSLCMRCFEDWNTKSKSCPFCRACLKKVNPSSLWLYTDDRDVVDMDTLTRENI RRLFMFISKLPLVVLHVVDLDIYEYRIK</aaseq>
Gene Sequence	<dnaseqindica>1840..2053#813..1035#649..715#292..528#ggggctcggattagagttggactcggcaacgcgacgagcaccaaaagcgagggaagaaaaccaatccgaatccggagagagaagcgcaggaggcaaaatccactccaattcaacccaatcacccgcggcgagccgagcgggcggcggagcacgacgacgacgaggtttaggtaggggttcttggcttgtcggcgtcggcggcggcggcggcggaggaggaggaggagtcagcgatgcggaggaggttccaggactccgtcaaggccctcgaggccgacatcgagcacgccaatgagctgtgagttcttcctcccttcgccggtcgcggctcgtggacggtggtggtggtggggggtttgcgtgtggttcatctctctcttttgcagggcgtcggagttcttgcgggattacgacggggcggtgatccagatgcggatggcgtacagcgccgtcgcgcacttcctcgtgcagtggatcgactgcaagctcgccggcgcgctcggcctcctcaagatcatgatctacaaggtctcgcccacacccccgcgccatcgccaattcgcagctgatttctcaccggctgggtaactaactaactaactaactaactaactaatcaccgggttcttgctggtaaatcgaatgcaggtgtacgccgatggcaccacggctctgccggagtgggagagggaggccagcatcaggcaattctacggtactactactcaatcgcaaccatctcctaacaacaagaacacgagcgattcaatttcatggttgtgatttctcaaattttttttgggtcgctgcaggtgtcatcttcccgtcgctgctccagctgccgagtgggataactgaattggacgacaggaagcagaggaggctgtgccttcagaagttcaggaaggtggaggagagggtctcggaggtggatttggagagggagctcgagtgcggcatctgcctcgaggtgaatgccaagattgtgctgcccgattgcgcgcactcgctgtgcatgagatgcttcgaggattggtaatttgccctcctccttcccctttaatttcccctctccttacattttcgcgcatgcgccaacacaaacatagagatattaggtactatcaattgttcatgttagaatcaaatatagctgttgcgtgctctaggagacaaaatttcgtttacctgaatgtggtgttcaagaaagagagaaaaagattacctttgccatatattggctggttgtgttcctcaggatgtaaagtcagttgtaaatctcctgcacttctgatagagtaccacaatgccctctccacagagtctttaaccatcttcatgcatcaagatgtagtccatccaatcaaacctgcatccagaatgactttatttataactgcagcaaacattctattgaactatgtcctgctcttgagcaggtaataacttattgatgtaaaagaagtaggcaaatagtcacaaaatgaactatatcatagatcggttgcatgatgctcctccaaaatacaaacctctgtatctggtaacaattctcaccttggttgaaggttacacacgcgcacacacagtctttgcatactctcttatgctgttagaaaactatgttgttctcagttcattgtatggttgacttcactttccttatagtcaatcctaagttgagagtgaatcattaaccactccttgtcagcaagcacaacaagatctgcaaccttgtccgtactcttcctacttgccgaggcgttcacttcgtctgatgcatgaatctgatcgaattccgtgggccatttaacgaaatttggcttgtcacatatgcaggaacaccaaatcaaagtcgtgccccttctgccgcgcctgcctcaagaaggtgaatccgagcagcctgtggttgtacaccgacgaccgcgatgttgtggatatggatacgttgactagggagaacattaggcgcctgttcatgttcataagtaagcttccacttgtagtgctccatgtggttgaccttgacatttacgagtaccgtatcaagtgaaactgtactcttttgttcataccggtgggtctctgtacatatcaaattcatcggtgctgatctgtgatagctcaacctgaggctgcaaattagcagagttgtttgtagctcaacgagttgataatatttttgtgaaaagaagatgctgaagtgtactcc</dnaseqindica>
External Link(s)	NCBI Gene:Os10g0445400, RefSeq:Os10g0445400