Difference between revisions of "Os07g0508500"

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Please input one-sentence summary here.
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The rice '''''Os07g0508500''''' was reported as '''''BRK1''''' in 2012 <ref name="ref1" /> by researchers from China.  
  
 
==Annotated Information==
 
==Annotated Information==
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[[File:108-Os07g0508500.png|right|thumb|327px|'''Figure 2.''' ''Meiotic Chromosome Behavior in the Wild Type and brk1-1 Mutant.<ref name="ref1" />.'']]
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===Gene Symbol===
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*'''''Os07g0508500''''' '''''<=>''''' '''''BRK1,OsBRK1'''''
 
===Function===
 
===Function===
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Please input function information here.
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* '''''BRK1''''' may be required for normal localization of Aurora kinase before the onset of metaphase I, which is responsible for correcting the merotelic attachment.
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* '''''BRK1''''' Is Essential for the Centromeric Localization of SGO1
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* '''''BRK1''''' Is Required for Phosphorylation of Rice H2A-T134 during Both Meiosis and Mitosis
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* Depletion of '''''BRK1''''' Reduces the Tension across Homologous Kinetochores during Metaphase I
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* '''''BRK1''''' Is Required for Correcting the Merotelic Attachment of Paired Sister Kinetochores during Metaphase I
 
===Expression===
 
===Expression===
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'''Plant Materials'''
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* During prophase I, chromosomal behavior in brk1-1 was quite similar to the wild type (Figures 2A and 2E). Whereas at the onset of anaphase I in brk1-1, although homologous chromosomes could separate equally, the two sister chromatids of each chromosome started to disassociate from each other (Figure 2F).
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* During telophase I in brk1-1, the sister chromatids of most chromosomes completely separated, and this precocious dissociation resulted in scattered chromatids that failed to line up on the equatorial plate during metaphase II (Figure 2G).  
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The blk1-1 mutant was isolated from the indica rice (Oryza sativa) variety
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* Consequently, sister chromatids were unequally separated at anaphase II in brk1-1, and the lagging chromosomes finally led to the formation of micronuclei during the tetrad stage (Figure 2H).
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Zhongxian 3037 following
 
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60
 
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Co;g-ray radiation. Seeds of the Tos17 insertion line NE7019, which we called blk1-2, were kindly provided by the
 
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Rice Genome Resource Center of the National Institute of Agrobiological
 
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Sciences. The other meiotic mutant, sgo1-1, was isolated in our lab previously (Wang et al., 2011). All plant material was cultivated in paddy fields.
 
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'''Molecular Cloning of BRK1 and Tos17 Insertion Site Mapping'''
 
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To fine map BRK1, STS markers (P1 to P5) were developed based on
 
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sequence differences between indica variety 9311 and japonica variety
 
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Zhonghua 11 according to data published on the National Center for
 
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Biotechnology Information website. All primer sequences are listed in
 
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Supplemental Table 1 online. Primer pair TosP (59-ATTGTTAGGTTGCAAGTTAGTTAAGA-39) and 7019P (59-TGTATCGCCAGCTTGATACG-
 
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39) was used to amplify the Tos17-inserted regions of brk1-2. The PCR
 
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products were cloned into the pMD18-T vector (TaKaRa) and sequenced.
 
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'''Cloning the Full-Length BRK1 cDNA'''
 
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Total RNA was extracted from rice panicle using TRIzol reagent (Invitrogen), and 3 mg RNA was reverse transcribed with the oligo-dT(18)
 
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primer using MMLV-RT (Invitrogen). Gene-specific primers forward (59-
 
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ATGGTTCTGCTCGAACGCAC-39) and reverse (59-GATGCGTATATGTCTCGGT-39) were designed to amplify the predicted coding region of
 
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BRK1. PCR products were cloned into the pMD18-T vector (TaKaRa) and
 
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sequenced. 39- and 59-RACE was performed according to the kit protocols (39-Full RACE Core Set and 59-Full RACE Core Set; TaKaRa). For
 
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39-RACE, the first and second PCRs were performed using 39RACE-F1
 
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(59-CTGCTCAACGCGACCTCCAA-39) and the 39 adaptor (59-AAGCAGTGGTATCAACGCAGA-39), and 39RACE-F2 (59-AGCCTCCGGATGTCCTTCCA-39) and the 39 adaptor, respectively. For 59-RACE, total RNA was
 
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reverse transcribed using the 59 phosphorylated primer 59RACE-RT (59-
 
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CCTTACTGAAGAGAAACAGT-39). First and second PCRs were performed using 59RACE-S1 (59-CTCAGGTCTATCAACAAGGCT-39) and
 
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59RACE-R1 (59-TGCTGCCTCGGGCACCACCAA-39), and 59RACE-S2
 
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(59-GATCGGCTCAAGATTTGTGT-39) and 59RACE-R2 (59-CGTGCGTTCGAGCAGAACCA-39), respectively. The products of 39-RACE-PCR and
 
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59-RACE-PCR were cloned and sequenced.
 
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'''Real-Time PCR Expression Assay'''
 
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Real-time PCR assay was conducted as described (Hong et al., 2010).
 
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Total RNA was extracted from the leaf, root, and panicle of Zhongxian
 
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3037 and panicles of brk1 mutants. Real-time PCR analysis was performed using the Bio-Rad CFX96 real-time PCR instrument and EvaGreen
 
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(BIOTIUM). Real-Time PCR was performed using primers BRKRT-F
 
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(59-CTGCTCAACGCGACCTCCAA-39) and BRKRT-R (59-GCTACGAAGTTACGGAAGGT-39) for BRK1 and UBQF (59-CAAGATGATCTGCCGCAAATGC-39) and UBQR (59-TTTAACCAGTCCATGAACCCG-39) for
 
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Ubiquitin. The real-time PCR results were analyzed using Bio-Rad CFX
 
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Manager analysis software. Each experiment had three replicates.
 
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'''Construction of BRK1 RNA Interference Cassette and
 
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Rice Transformation'''
 
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A 310-bp fragment of the BRK1 cDNA sequence was amplified with the
 
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primer pair RNAiF, 59-ATTCTCGAGGCCAGTGTCGATGGTAATACA-39
 
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(adding an XhoI site), and RNAiR, 59-ACGAGATCTCAACGCTGTGCAGTGTTTCCA-39 (adding a BglII site). The fragment was cloned into the
 
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BamHI-SalI and BglII-XhoI sites of the pUCRNAi vector. Subsequently,
 
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the stem-loop fragment was cloned into the pCAMBIA2300-Actin vector.
 
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Then the RNAi construct was transformed into an Agrobacterium tumefaciens strain, which was used for further rice transformation.
 
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'''Antibody Production and Protein Gel Blot Analysis'''
 
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Antibodies were produced as described (Wang et al., 2010). To generate
 
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the antibody against BRK1, a 330-bp fragment of the BRK1 cDNA (amino
 
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acids 396 to 505) was amplified with primers BRKAbF, 59-GTGAATTCCCGAAATCACAGACGAAACCTT-39 (adding an EcoRI site), and
 
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BRKAbR, 59-AGTCTCGAGTCCACAGCTCCACGTTCCAGT-39 (adding an
 
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XhoI site). The fragment was cloned into the expression vector pGEX 4T-2.
 
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The BRKAb-GST peptides were purified using Glutathione Sepharose 4B
 
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(GE). Polyclonal antibodies against BRK1 were raised in rabbit and mouse.
 
  
 
===Evolution===
 
===Evolution===
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Please input evolution information here.
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* BRK1 Localizes to Kinetochores during Both Meiosis and Mitosis
  
 
You can also add sub-section(s) at will.
 
You can also add sub-section(s) at will.
  
 
==Labs working on this gene==
 
==Labs working on this gene==
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Please input related labs here.
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* State Key Laboratory of Plant Genomics and Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
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* College of Plant Protection,Yunnan Agricultural University, Kunming 650201, China
 
==References==
 
==References==
 
<references>
 
<references>
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<ref name="ref1">BRK1, a Bub1-Related Kinase, Is Essential for Generating Proper Tension between Homologous Kinetochores at Metaphase I of Rice Meiosis</ref>
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* <ref name="ref1">
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Wang M, Tang D, Luo Q, Jin Y, Shen Y, Wang K, Cheng Z. BRK1, a Bub1-related
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kinase, is essential for generating proper tension between homologous
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kinetochores at metaphase I of rice meiosis. Plant Cell. 2012 Dec;24(12):4961-73.
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doi: 10.1105/tpc.112.105874. Epub 2012 Dec 14. PubMed PMID: 23243128; PubMed
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Central PMCID: PMC3556969.
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</ref>
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</references>
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==Structured Information==

Latest revision as of 15:50, 17 October 2016

The rice Os07g0508500 was reported as BRK1 in 2012 [1] by researchers from China.

Annotated Information

Figure 2. Meiotic Chromosome Behavior in the Wild Type and brk1-1 Mutant.[1].

Gene Symbol

  • Os07g0508500 <=> BRK1,OsBRK1

Function

  • BRK1 may be required for normal localization of Aurora kinase before the onset of metaphase I, which is responsible for correcting the merotelic attachment.
  • BRK1 Is Essential for the Centromeric Localization of SGO1
  • BRK1 Is Required for Phosphorylation of Rice H2A-T134 during Both Meiosis and Mitosis
  • Depletion of BRK1 Reduces the Tension across Homologous Kinetochores during Metaphase I
  • BRK1 Is Required for Correcting the Merotelic Attachment of Paired Sister Kinetochores during Metaphase I

Expression

  • During prophase I, chromosomal behavior in brk1-1 was quite similar to the wild type (Figures 2A and 2E). Whereas at the onset of anaphase I in brk1-1, although homologous chromosomes could separate equally, the two sister chromatids of each chromosome started to disassociate from each other (Figure 2F).
  • During telophase I in brk1-1, the sister chromatids of most chromosomes completely separated, and this precocious dissociation resulted in scattered chromatids that failed to line up on the equatorial plate during metaphase II (Figure 2G).
  • Consequently, sister chromatids were unequally separated at anaphase II in brk1-1, and the lagging chromosomes finally led to the formation of micronuclei during the tetrad stage (Figure 2H).

Evolution

  • BRK1 Localizes to Kinetochores during Both Meiosis and Mitosis

You can also add sub-section(s) at will.

Labs working on this gene

  • State Key Laboratory of Plant Genomics and Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
  • College of Plant Protection,Yunnan Agricultural University, Kunming 650201, China

References

  1. ↑ 1.0 1.1 Wang M, Tang D, Luo Q, Jin Y, Shen Y, Wang K, Cheng Z. BRK1, a Bub1-related kinase, is essential for generating proper tension between homologous kinetochores at metaphase I of rice meiosis. Plant Cell. 2012 Dec;24(12):4961-73. doi: 10.1105/tpc.112.105874. Epub 2012 Dec 14. PubMed PMID: 23243128; PubMed Central PMCID: PMC3556969.

Structured Information

Retrieved from "https://ngdc.cncb.ac.cn/ricewiki/index.php?title=Os07g0508500&oldid=274877"