Difference between revisions of "Os06g0472000"
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| − | + | ''OsMSRB1.1'' is a mumber of MSRB, which belongs to methionine sulfoxide reductases (MSRs)<ref name="ref1"/><ref name="ref2"/>. | |
==Annotated Information== | ==Annotated Information== | ||
===Function=== | ===Function=== | ||
| − | + | *Expression analysis showed that both ''OsMSRA4.1'' and ''OsMSRB1.1'' are constitutively expressed in all organs and can be induced by various stress conditions. Overexpression of either ''OsMSRA4.1'' or ''OsMSRB1.1'' in yeast enhanced cellular resistance to oxidative stress. In addition, OsMSRA4.1-overexpressing transgenic rice plants also showed enhanced viability under salt treatment<ref name="ref1"/>. | |
| + | *Cloning of ''OsMSRB1.1'' cDNAs: Total RNA from rice leaves was extracted using the guanidinium isocyanate/acidic phenol method as described by ''Chomczynski and Sacchi''. cDNA synthesis and RTPCR were performed using a kit. The ''OsMSRB1.1'' gene-specific primers were 5'-AATGGCCATGCGGCAATAC-3' and 5'-CACAGGTCTACTGGGTCTTCTTCAG-3'. The ''OsMSRB1.1'' cDNAs was cloned into the yeast expression vector p181AINE and the constructs were transformed into yeast using the lithium acetate method<ref name="ref1"/>. | ||
| + | *Analysis of the EST sequences suggests that ''OsMSRB1'' transcript is probably alternatively spliced, leading to two distinct proteins (OsMSRB1.1 and 1.2), which differ only by few residues located in the C-terminal part<ref name="ref2"/>. | ||
===Expression=== | ===Expression=== | ||
| − | + | *''OsMSRB1.1'' partially enhanced the tolerance of the yeast double mutant in the absence of H<sub>2</sub>O<sub>2</sub>. The transgenic yeast exhibited reduced sensitivity to H<sub>2</sub>O<sub>2</sub> compared with the double mutant when treated with 2 mM H<sub>2</sub>O<sub>2</sub>. In addition, overexpression of ''OsMSRB1.1'' in wild-type yeast also accelerated the growth rate under oxidative stress. | |
| + | *To better understand the functions, the expression patterns of ''OsMSRA4.1'' and ''OsMSRB1.1'' were examined by RT-PCR in difierent tissues. ''OsMSRB1.1'' was found to be preferentially expressed in leaves, Xowers, and callus, with weak expression in roots and stems. | ||
| + | *To investigate the transcription inducibility of OsMSRs, Northern blot analysis was performed under various stress conditions. | ||
| + | ''OsMSRB1.1'' expression was up-regulated by various abiotic stresses including salt, mannitol, MV, cold and ABA. But under high temperature treatment, the expression level of ''OsMSRB1.1'' decreased during treatment. | ||
| + | *''OsMSRA4.1'' and ''OsMSRB1.1'' were able to reduce both the free MetSO and protein-bound-like MetSO substrates in the presence of | ||
| + | the DTT reducing system. In addition, the proteins showed higher catalytic activities with dabsyl-MetSO than free MetSO as a substrate. | ||
| + | |||
| + | ===Subcellular localization=== | ||
| + | Subcellular localization and in vitro activity assay revealed that both OsMSR proteins are targeted to the chloroplast and have MSR activity. Based on the prediction software (TargetP), ''OsMSRA4'', ''OsMSRB1'', and ''OsMSRB3'' all have N-terminal extensions which are predicted to be chloroplast transit peptides. ''OsMSRA5'' is predicted to be localized to a secretory pathway. ''OsMSRA2.1'', ''OsMSRA2.2'', and ''OsMSRB5'' are possibly restricted to cytosol (Table 1), but the exact subcellular localization remains to be experimentally investigated<ref name="ref1"/>. | ||
===Evolution=== | ===Evolution=== | ||
| − | + | [[File: MSR gene family.jpg|left|thumb|300px|'''Table 1.'''''MSR gene family in rice.(from reference <ref name="ref1"/>).'']] | |
| + | [[File: MSR Phylogenetic tree.jpg|right|thumb|250px|'''Figure 1.'''''Phylogenetic tree of MSRAs and MSRBs from Arabidopsis thaliana, Populus trichocarpa and Oryza sativa.(from reference <ref name="ref2"/>).'']] | ||
| + | *Using the blastP search engine in the NCBI database, seven MSR genes were found in rice genome (Table 1). To avoid confusion, | ||
| + | Sequence alignment revealed that ''OsMSRA2.1'', ''OsMSRA2.2'', ''OsMSRA4'', and ''OsMSRA5'' belong to the MSRA, and ''OsMSRB1'', ''OsMSRB3'', and ''OsMSRB5'' belong to the MSRB (Table 1). Accession numbers and chromosome locations of rice MSR gene family are listed in Table 1. Two transcripts (''OsMSRA4.1/2'' and ''OsMSRB1.1/2'') with diVerent lengths for OsMSRA4 and OsMSRB1 were found, respectively, probably resulted from alternative splicing<ref name="ref1"/>. | ||
| + | *Multiple alignment of ''OsMSRA'' sequences revealed that high similarity was found among ''OsMSRA2.1'', ''OsMSRA2.2'', and ''OsMSRA4''. Three of the OsMSRA proteins contain one highly conserved cysteine residue housed in the GCFWG motif, while ''OsMSRA5'' possesses a serine residue instead of this cysteine residue. OsMSRB sequences are also highly conserved and all OsMSRBs have four additional conserved cysteine residues that are organized in two CXXC motifs (two cysteines separated by two amino acid residues) which were found to coordinate structural Zinc<ref name="ref1"/>. | ||
| + | *''OsMSRA4.1'' and ''OsMSRB1.1'' orthologs in Arabidopsis：''AtMSRA4'' and ''AtMSRB1''<ref name="ref3"/><ref name="ref4"/>. | ||
| + | *Based on sequence alignments, on construction of unrooted phylogenetic trees and on some biochemical results, two MSRA subgroups can be distinguished. They differ essentially in the number and in the position of the cysteines involved in catalysis and enzyme | ||
| + | regeneration, but also in the subcellular localization. ''MSRA5'' isoforms constitute an independent subgroup, while other proteins, either cytosolic or chloroplastic, are grouped into the same clad (Fig. 1). When looking at sequence homology, the overall identities range from 55 to 66% for MSRA5 proteins, from 55 to 93% for MSRA1-4, but only from 21 to 34% between ''MSRA5'' proteins and other MSRAs.From the phylogenetic tree shown in Fig. 2, it appears that the chloroplastic MSRB1 isoforms constitute a distinct subgroup, while all other MSRB are grouped together<ref name="ref2"/>. | ||
| − | + | ===Knowledge Extension=== | |
| + | In rice genome, MSR is encoded by a multigene family with at least seven members. The relatively large number of MSR genes in plants is particularly signiWcant. In ''Arabidopsis'', the number of genes encoding for MSRs is even larger with 14 members, when compared to 9 in poplar and 7 in rice<ref name="ref5"/> whereas generally mammals, yeast, and E. coli possess 2–4 MSR genes. | ||
==Labs working on this gene== | ==Labs working on this gene== | ||
| − | + | *State Key Laboratory of Plant Genomics, National Centre for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences (CAS), 100101 Beijing, China | |
| + | *Graduate School of the Chinese Academy of Sciences, 100039 Beijing, China | ||
==References== | ==References== | ||
| − | + | <references> | |
| + | * <ref name="ref1"> | ||
| + | Guo X, Wu Y, Wang Y, et al. OsMSRA4. 1 and OsMSRB1. 1, two rice plastidial methionine sulfoxide reductases, are involved in abiotic stress responses[J]. Planta, 2009, 230(1): 227-238. | ||
| + | </ref> | ||
| + | * <ref name="ref2"> | ||
| + | Rouhier N, Dos Santos C V, Tarrago L, et al. Plant methionine sulfoxide reductase A and B multigenic families[J]. Photosynthesis research, 2006, 89(2-3): 247-262. | ||
| + | </ref> | ||
| + | * <ref name="ref3"> | ||
| + | Romero H M, Berlett B S, Jensen P J, et al. Investigations into the role of the plastidial peptide methionine sulfoxide reductase in response to oxidative stress in Arabidopsis[J]. Plant physiology, 2004, 136(3): 3784-3794. | ||
| + | </ref> | ||
| + | * <ref name="ref4"> | ||
| + | Dos Santos C V, Cuiné S, Rouhier N, et al. The Arabidopsis plastidic methionine sulfoxide reductase B proteins. Sequence and activity characteristics, comparison of the expression with plastidic methionine sulfoxide reductase A, and induction by photooxidative stress[J]. Plant physiology, 2005, 138(2): 909-922. | ||
| + | </ref> | ||
| + | * <ref name="ref5"> | ||
| + | Rouhier N, Dos Santos C V, Tarrago L, et al. Plant methionine sulfoxide reductase A and B multigenic families[J]. Photosynthesis research, 2006, 89(2-3): 247-262. | ||
| + | </ref> | ||
| + | </references> | ||
==Structured Information== | ==Structured Information== | ||
Revision as of 14:59, 3 January 2015
OsMSRB1.1 is a mumber of MSRB, which belongs to methionine sulfoxide reductases (MSRs)[1][2].
Contents
Annotated Information
Function
- Expression analysis showed that both OsMSRA4.1 and OsMSRB1.1 are constitutively expressed in all organs and can be induced by various stress conditions. Overexpression of either OsMSRA4.1 or OsMSRB1.1 in yeast enhanced cellular resistance to oxidative stress. In addition, OsMSRA4.1-overexpressing transgenic rice plants also showed enhanced viability under salt treatment[1].
- Cloning of OsMSRB1.1 cDNAs: Total RNA from rice leaves was extracted using the guanidinium isocyanate/acidic phenol method as described by Chomczynski and Sacchi. cDNA synthesis and RTPCR were performed using a kit. The OsMSRB1.1 gene-specific primers were 5'-AATGGCCATGCGGCAATAC-3' and 5'-CACAGGTCTACTGGGTCTTCTTCAG-3'. The OsMSRB1.1 cDNAs was cloned into the yeast expression vector p181AINE and the constructs were transformed into yeast using the lithium acetate method[1].
- Analysis of the EST sequences suggests that OsMSRB1 transcript is probably alternatively spliced, leading to two distinct proteins (OsMSRB1.1 and 1.2), which differ only by few residues located in the C-terminal part[2].
Expression
- OsMSRB1.1 partially enhanced the tolerance of the yeast double mutant in the absence of H2O2. The transgenic yeast exhibited reduced sensitivity to H2O2 compared with the double mutant when treated with 2 mM H2O2. In addition, overexpression of OsMSRB1.1 in wild-type yeast also accelerated the growth rate under oxidative stress.
- To better understand the functions, the expression patterns of OsMSRA4.1 and OsMSRB1.1 were examined by RT-PCR in difierent tissues. OsMSRB1.1 was found to be preferentially expressed in leaves, Xowers, and callus, with weak expression in roots and stems.
- To investigate the transcription inducibility of OsMSRs, Northern blot analysis was performed under various stress conditions.
OsMSRB1.1 expression was up-regulated by various abiotic stresses including salt, mannitol, MV, cold and ABA. But under high temperature treatment, the expression level of OsMSRB1.1 decreased during treatment.
- OsMSRA4.1 and OsMSRB1.1 were able to reduce both the free MetSO and protein-bound-like MetSO substrates in the presence of
the DTT reducing system. In addition, the proteins showed higher catalytic activities with dabsyl-MetSO than free MetSO as a substrate.
Subcellular localization
Subcellular localization and in vitro activity assay revealed that both OsMSR proteins are targeted to the chloroplast and have MSR activity. Based on the prediction software (TargetP), OsMSRA4, OsMSRB1, and OsMSRB3 all have N-terminal extensions which are predicted to be chloroplast transit peptides. OsMSRA5 is predicted to be localized to a secretory pathway. OsMSRA2.1, OsMSRA2.2, and OsMSRB5 are possibly restricted to cytosol (Table 1), but the exact subcellular localization remains to be experimentally investigated[1].
Evolution
- Using the blastP search engine in the NCBI database, seven MSR genes were found in rice genome (Table 1). To avoid confusion,
Sequence alignment revealed that OsMSRA2.1, OsMSRA2.2, OsMSRA4, and OsMSRA5 belong to the MSRA, and OsMSRB1, OsMSRB3, and OsMSRB5 belong to the MSRB (Table 1). Accession numbers and chromosome locations of rice MSR gene family are listed in Table 1. Two transcripts (OsMSRA4.1/2 and OsMSRB1.1/2) with diVerent lengths for OsMSRA4 and OsMSRB1 were found, respectively, probably resulted from alternative splicing[1].
- Multiple alignment of OsMSRA sequences revealed that high similarity was found among OsMSRA2.1, OsMSRA2.2, and OsMSRA4. Three of the OsMSRA proteins contain one highly conserved cysteine residue housed in the GCFWG motif, while OsMSRA5 possesses a serine residue instead of this cysteine residue. OsMSRB sequences are also highly conserved and all OsMSRBs have four additional conserved cysteine residues that are organized in two CXXC motifs (two cysteines separated by two amino acid residues) which were found to coordinate structural Zinc[1].
- OsMSRA4.1 and OsMSRB1.1 orthologs in Arabidopsis：AtMSRA4 and AtMSRB1[3][4].
- Based on sequence alignments, on construction of unrooted phylogenetic trees and on some biochemical results, two MSRA subgroups can be distinguished. They differ essentially in the number and in the position of the cysteines involved in catalysis and enzyme
regeneration, but also in the subcellular localization. MSRA5 isoforms constitute an independent subgroup, while other proteins, either cytosolic or chloroplastic, are grouped into the same clad (Fig. 1). When looking at sequence homology, the overall identities range from 55 to 66% for MSRA5 proteins, from 55 to 93% for MSRA1-4, but only from 21 to 34% between MSRA5 proteins and other MSRAs.From the phylogenetic tree shown in Fig. 2, it appears that the chloroplastic MSRB1 isoforms constitute a distinct subgroup, while all other MSRB are grouped together[2].
Knowledge Extension
In rice genome, MSR is encoded by a multigene family with at least seven members. The relatively large number of MSR genes in plants is particularly signiWcant. In Arabidopsis, the number of genes encoding for MSRs is even larger with 14 members, when compared to 9 in poplar and 7 in rice[5] whereas generally mammals, yeast, and E. coli possess 2–4 MSR genes.
Labs working on this gene
- State Key Laboratory of Plant Genomics, National Centre for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences (CAS), 100101 Beijing, China
- Graduate School of the Chinese Academy of Sciences, 100039 Beijing, China
References
- ↑ 1.0 1.1 1.2 1.3 1.4 1.5 1.6 Guo X, Wu Y, Wang Y, et al. OsMSRA4. 1 and OsMSRB1. 1, two rice plastidial methionine sulfoxide reductases, are involved in abiotic stress responses[J]. Planta, 2009, 230(1): 227-238.
- ↑ 2.0 2.1 2.2 2.3 Rouhier N, Dos Santos C V, Tarrago L, et al. Plant methionine sulfoxide reductase A and B multigenic families[J]. Photosynthesis research, 2006, 89(2-3): 247-262.
- ↑ Romero H M, Berlett B S, Jensen P J, et al. Investigations into the role of the plastidial peptide methionine sulfoxide reductase in response to oxidative stress in Arabidopsis[J]. Plant physiology, 2004, 136(3): 3784-3794.
- ↑ Dos Santos C V, Cuiné S, Rouhier N, et al. The Arabidopsis plastidic methionine sulfoxide reductase B proteins. Sequence and activity characteristics, comparison of the expression with plastidic methionine sulfoxide reductase A, and induction by photooxidative stress[J]. Plant physiology, 2005, 138(2): 909-922.
- ↑ Rouhier N, Dos Santos C V, Tarrago L, et al. Plant methionine sulfoxide reductase A and B multigenic families[J]. Photosynthesis research, 2006, 89(2-3): 247-262.
Structured Information
| Gene Name |
Os06g0472000 |
|---|---|
| Description |
Mss4-like domain containing protein |
| Version |
NM_001064155.1 GI:115468041 GeneID:4341018 |
| Length |
2685 bp |
| Definition |
Oryza sativa Japonica Group Os06g0472000, 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 6:16595548..16598232 |
| Sequence Coding Region |
16595765..16595795,16596503..16596700,16596842..16596907,16596995..16597103,16597998..16598232 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008399:16595548..16598232 source=RiceChromosome06 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008399:16595548..16598232 source=RiceChromosome06 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atgcggcaatacgcggctgctaccgctgcctcctccagtttcagagcacgtccacgggcgcgcccctcctgcctcccagccgccgccctgcccttggcgccttgctgtggtgtggcgtggagccgtgctagctacaggcgagcctccgttcgtgccatgggtgccgcttcatcgtcttcgtcgtcgtcgtcgtcgtctccgtcgccgcagggtcaagcccaagcccaagcccaaggtaaaccgaactacagtacatctctgactgatgaggagtggaggaagcgcctgacaaaagatcagtattacattactcggcagaagggcacagaaagagcatttactggggaatactggaacaccaaaaccccgggcatctaccattgtgtctgctgtgacacccctctttttgagtcatcgaccaaatttgatagtggtactgggtggccgtcatattatcaacccattggagataatgtaaagtgcaagcttgatatgtccatcatattcatgcctcggactgaggtgctgtgtgctgtctgtgacgctcatctggggcacgtgtttgatgatgggccacgaccaacagggaaaagatactgtatcaatagcgcatctctcaagctgaagaagacccagtag</cdnaseq> |
| Protein Sequence |
<aaseq>MRQYAAATAASSSFRARPRARPSCLPAAALPLAPCCGVAWSRAS YRRASVRAMGAASSSSSSSSSSPSPQGQAQAQAQGKPNYSTSLTDEEWRKRLTKDQYY ITRQKGTERAFTGEYWNTKTPGIYHCVCCDTPLFESSTKFDSGTGWPSYYQPIGDNVK CKLDMSIIFMPRTEVLCAVCDAHLGHVFDDGPRPTGKRYCINSASLKLKKTQ</aaseq> |
| Gene Sequence |
<dnaseqindica>2438..2468#1533..1730#1326..1391#1130..1238#1..235#atgcggcaatacgcggctgctaccgctgcctcctccagtttcagagcacgtccacgggcgcgcccctcctgcctcccagccgccgccctgcccttggcgccttgctgtggtgtggcgtggagccgtgctagctacaggcgagcctccgttcgtgccatgggtgccgcttcatcgtcttcgtcgtcgtcgtcgtcgtctccgtcgccgcagggtcaagcccaagcccaagcccaaggtgccgtgtggtgcttcaccttctaacttctttacctacctagtgggagcccgttgttgcctgcttcctttggatgcaagggaattattcgcgagtggaaatagaaattgaattaatggtgattagtttgtatgtgcattcgatgcaaacaaggagggagtttgattgatcttaaaagatctgctgtaggccgtttctatgtctaattctttccgcatttaagaacattttagaaaattactagaccgtaattcgggagtgaagcccctattgcccgctcaagtttgcaactaagacatttttataagttagctacatagatcctgagtggttttagctattttcattcaaacagttatatgttactcacaagtgtattcaccagtcaatagtgcggacattctttataggtgtgaatttcggactgtgtacctcaaagacaaaaaaaaaaaaagaaagaaagaaagaaaagaattctgtgttttagttcagagtacagaacatatagtataaacacacatcagttacaaatctaaggtaatcttaagagaattgtgcattcttgctatatcctggtagagccttacaagcatttgtatgcagtgagtcactgagctgactagtatcgattttgtctatgccagcagctccttcttttatagtttgttagaacaattgatcttcagggttcacaatgtggagcaaatctttaagagaataatgcattgatgttctgctttcttatggagcaaatgcttagaatgccgcctggcttgaagatatttttttatcatacgctcttaactctgaagtactccagatctgtatgtagccaggttccaacaattgtttcaattctcactaatagccaaatattcatttcttctgaggcaggtaaaccgaactacagtacatctctgactgatgaggagtggaggaagcgcctgacaaaagatcagtattacattactcggcagaagggcacagaaagagcatttactgggtacgattgctttacactctaatccaagtataatgtgtagttgcacaccatgtcattttggtcttgtgttttttgttatgtaatcagggaatactggaacaccaaaaccccgggcatctaccattgtgtctgctgtgacacccctctttttgagtaagcactcctcctgattgtcatgtcagagcagaagattgttctttcatataagctagagatagtagtttcgatttgcaccaaaacaatagtattatagtctgctcattattctgcattactgctttctggtggtcttaggtcatcgaccaaatttgatagtggtactgggtggccgtcatattatcaacccattggagataatgtaaagtgcaagcttgatatgtccatcatattcatgcctcggactgaggtgctgtgtgctgtctgtgacgctcatctggggcacgtgtttgatgatgggccacgaccaacagggaaaagatactgtatcaataggtatatgtgagattttcttcacttgttatcaattgttcataggcctcatttgaaatctgaacagaaacattgtctcccatatctacgaaattattttttttgaggggtatctacgaaattaactgtttgcttaattttcccactaacatttttcccattgttcaactccggtaagtcccagattagtttctaagtaaacttccgtgcatatatacatagtagtaccatagctaatatctgtctcatatgtgttgcattgcaccgaaggacctttctatacatgttcttttggttgatacatgactgcagagttgcagcttgccaacagtggatccaaaggcccacacgtgtataggtgccaattacaactgaatgcactctgaccaaatcttcaaaaaggaaaaaaaaaactgtccttacgtataccactgcttgaaagagaatcacattttcttttacaacagatgaaatattttactagggcatggttgccatgaattagtttataaaatttagtagcatccagtgaaatagaaccgccttttaatttatatttcctagataatggtaatttcactaggcaaaagaaagcattcagcttctgcacaaagtgcatataaccaacaacaccacgacacacacaaaagaaggaaccgtctttagtcgatgagattgtttgctcactgttttatccttcttacttgcagcgcatctctcaagctgaagaagacccagtagacctgtgaagattatggttaccatgtacctacacggctacactatccgactagttataagggatagattatgtatatagaatatgtaaagaattaagacttgggtattatgtttacttgtgtatcaaggaaatgacgtgtagtcctagttcggtaagattgtaaagtagttgtaggagatttaaaccatattaacttaaatctctatcttataaacc</dnaseqindica> |
| External Link(s) |
