BRK1, a Bub1-Related Kinase, Is Essential for Generating Proper Tension between Homologous Kinetochores at Metaphase I of Rice Meiosis

Wang, Mo; Tang, Ding; Luo, Qiong; Jin, Y.; Shen, Yi; Wang, Kejian; Cheng, Zhukuan

doi:10.1105/tpc.112.105874

Cited by 48 publications

(35 citation statements)

References 60 publications

(73 reference statements)

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“…Brk1 encodes a spindle checkpoint kinase acting during rice meiosis. A mutant allele of this gene was reported to be male sterile (Wang et al, 2012), which is consistent with our observation.…”

Section: Resultssupporting

confidence: 93%

Simultaneous Identification of Multiple Causal Mutations in Rice

Wang

Chen²,

Lü³

et al. 2017

Front. Plant Sci.

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Next-generation sequencing technologies (NGST) are being used to discover causal mutations in ethyl methanesulfonate (EMS)-mutagenized plant populations. However, the published protocols often deliver too many candidate sites and sometimes fail to find the mutant gene of interest. Accurate identification of the causal mutation from massive background polymorphisms and sequencing deficiencies remains challenging. Here we describe a NGST-based method, named SIMM, that can simultaneously identify the causal mutations in multiple independent mutants. Multiple rice mutants derived from the same parental line were back-crossed, and for each mutant, the derived F2 individuals of the recessive mutant phenotype were pooled and sequenced. The resulting sequences were aligned to the Nipponbare reference genome, and single nucleotide polymorphisms (SNPs) were subsequently compared among the mutants. Allele index (AI) and Euclidean distance (ED) were incorporated into the analysis to reduce noises caused by background polymorphisms and re-sequencing errors. Corrections of sequence bias against GC-and AT-rich sequences in the candidate region were conducted when necessary. Using this method, we successfully identified seven new mutant alleles from Huanghuazhan (HHZ), an elite indica rice cultivar in China. All mutant alleles were validated by phenotype association assay. A pipeline based on Perl scripts for SIMM is publicly available at https://sourceforge.net/projects/simm/.Keywords: SIMM, next-generation sequencing technology, single nucleotide polymorphism, SNP index, allele index, Euclidean distance, sequence correction INTRODUCTIONForward genetics based on EMS mutagenesis and cloning of mutant genes are of fundamental importance to the understanding of plant gene functions. Traditional map-based cloning approach has been widely used for cloning of mutant plant genes. It involves construction of mapping population(s) by crossing the mutant plant with wild-type plant of extensive genomic polymorphisms (Lukowitz et al., 2000). DNA sequence polymorphisms, such as simple sequence repeats, SNPs, and short insertions or deletions, are often used as markers for determination of parental chromosomal locations in the mapping population (Peters et al., 2003). By identifying markers that are tightly associated with the mutant phenotype, chromosomal region harboring Like map-based cloning, SHOREmap and NGM both involve crossing of the mutant with a remotely related wild-type plant; the derived F2 plants of the mutant phenotype are bulksequenced. Both methods have to deal with the numerous background polymorphisms between the two crossing accessions, which render it difficult to determine causal mutations. Thus far, SHOREmap, NGM, and other related methods have been successful only for cloning of mutant genes in Arabidopsis (Schneeberger et al., 2009;Austin et al., 2011;Uchida et al., 2011).MutMap (Abe et al., 2012) takes a different approach by back-crossing the mutant with the wild-type parent, and 20-30 derived F2 individuals of th...

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Section: Resultssupporting

confidence: 93%

Simultaneous Identification of Multiple Causal Mutations in Rice

Wang

Chen²,

Lü³

et al. 2017

Front. Plant Sci.

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“…S1). BRK1 displays 49% and 61% identity over its entire length with the recently characterized BRK1 from monocots ( Oryza sativa ) and dicots ( Vitis vinifera ), respectively (Karpov et al ., ; Wang et al ., ) (Fig. S2a).…”

Section: Resultsmentioning

confidence: 99%

Retracted: Three BUB1 and BUBR1/MAD3‐related spindle assembly checkpoint proteins are required for accurate mitosis in Arabidopsis

et al. 2014

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SummaryThe spindle assembly checkpoint (SAC) is a refined surveillance mechanism which ensures that chromosomes undergoing mitosis do not segregate until they are properly attached to the spindle microtubules (MT). The SAC has been extensively studied in metazoans and yeast, but little is known about its role in plants.We identified proteins interacting with a MT-associated protein MAP65-3, which plays a critical role in organising mitotic MT arrays, and carried out a functional analysis of previously and newly identified SAC components.We show that Arabidopsis SAC proteins BUB3.1, MAD2, BUBR1/MAD3s and BRK1 interact with each other and with MAP65-3. We found that two BUBR1/MAD3s interacted specifically at centromeres. When stably expressed in Arabidopsis, BRK1 localised to the kinetochores during all stages of the mitotic cell cycle. Early in mitosis, BUB3.1 and BUBR1/MAD3.1 localise to the mitotic spindle, where MAP65-3 organises spindle MTs. A double-knockout mad3.1 mad3.2 mutant presented spindle MT abnormalities, chromosome misalignments on the metaphase plate and the production of lagging chromosomes and micronuclei during mitosis.We conclude that BRK1 and BUBR1/MAD3-related proteins play a key role in ensuring faithful chromosome segregation during mitosis and that their interaction with MAP65-3 may be important for the regulation of MT-chromosome attachment.

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“…However, the function of SAC-related proteins in mitosis has not yet been tested in plants, and several SAC proteins have been found to be involved in plant meiosis, such as MPS1, Aurora kinases, AtPCH2, and CDC20.1 in Arabidopsis, as well as CRC1 and BRK1 in rice (12,37). In rice brk1-1 meiocytes, the spindle length was slightly elongated (38). In Arabidopsis cdc20.1, meiocytes exhibited atypical spindle morphology and had a longer average spindle length at metaphase I (37).…”

Section: Discussionmentioning

confidence: 99%

“…Highly conserved among eukaryotes, SAC proteins regulate chromosome segregation by sequestering CDC20 and promote the transition from metaphase to anaphase (36). Previous findings also reported that, in rice SAC protein Bub1-related kinase 1 (brk1) and Arabidopsis cell division cycle 20.1 (cdc20.1) mutants, the spindle lengths were a little bit longer than in the WT at metaphase I (37,38). Because p31 comet acts as an interacting partner of SAC effector molecule Mad2 in human (29), we thus investigated whether the loss of P31 comet is associated with the aberrant spindle assembly.…”

Section: Organization Of the Axial Elements And Central Elements Inmentioning

confidence: 98%

P31 ^comet , a member of the synaptonemal complex, participates in meiotic DSB formation in rice

Tang

Shen

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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The human mitotic arrest-deficient 2 (Mad2) binding protein p31 comet participates in the spindle checkpoint and coordinates cell cycle events in mitosis although its function in meiosis remains unknown in all organisms. Here, we reveal P31 comet as a synaptonemal complex (SC) protein in rice (Oryza sativa L.). In p31 comet , homologous pairing and synapsis are eliminated, leading to the homologous nondisjunction and complete sterility. The failure in loading of histone H2AX phosphorylation (γH2AX) in p31 comet , together with the suppressed chromosome fragmentation in rice completion of meiotic recombination 1 (com1) p31 comet and radiation sensitive 51c (rad51c) p31 comet double mutants, indicates that P31 comet plays an essential role in double-strand break (DSB) formation. Interestingly, the dynamic colocalization pattern between P31 comet and ZEP1 (a transverse filament protein of SC) by immunostaining, as well as the interaction between P31 comet and CENTRAL REGION COMPONENT 1 (CRC1) in yeast two-hybrid assays, suggests possible involvement of P31 comet in SC installation. Together, these data indicate that P31 comet plays a key role in DSB formation and SC installation, mainly through its cooperation with CRC1.uring meiosis, homologous recombination plays a crucial role in ensuring faithful and precise chromosome segregation, as well as in diversifying genetic information. Meiotic recombination is induced by the programmed formation of DNA double-strand breaks (DSBs), catalyzed by Spo11, a functional homolog of subunit A of an archaeal topoisomerase (TopoVIA) (1). In Saccharomyces cerevisiae, nine associated proteins, including meiotic recombination 11 (Mre11), radiation sensitive 50 (Rad50), X-ray sensitive 2 (Xrs2), superkiller 8 (Ski8), Rec102, Rec104, Rec114, meiosis-specific 4 (Mei4), and Mer2, are required for facilitating Spo11's catalytic reaction (2). In Schizosaccharomyces pombe, seven DSB formation-related proteins were identified (3). Rec6 has been recognized as an essential component for ensuring the meiotic DSB formation, and it forms a complex with Rec12 and Rec14, the orthologs of Spo11 and Ski8, respectively (4). Furthermore, the Mde2 protein, which lacks an ortholog in S. cerevisiae, is also required for DSB formation (5). Nevertheless, Rad50, Mre11, and Xrs2 orthologs are dispensable for DSB formation in S. pombe, as well as in mice and plants, but they are required for meiotic DSB processing (6). In mice, except for the conserved protein Spo11, Mei4 and Rec114 were also identified as key components involved in DSB formation (7). In addition, Mei1 was recognized as a recombination related factor, without an ortholog in S. cerevisiae or S. pombe (8). Recently, mouse TOPOVIBL protein was identified to form the complex with Spo11 and take part in meiotic DSB formation (9).In plants, the Arabidopsis AtSPO11-1, AtSPO11-2, and AtSPO11-3 share sequence similarity with other Spo11/topo VIA proteins. AtSPO11-1 and AtSPO11-2, as subunits of a heterodimer, are essential for DSB formation whereas A...

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BRK1, a Bub1-Related Kinase, Is Essential for Generating Proper Tension between Homologous Kinetochores at Metaphase I of Rice Meiosis

Cited by 48 publications

References 60 publications

Simultaneous Identification of Multiple Causal Mutations in Rice

Simultaneous Identification of Multiple Causal Mutations in Rice

Retracted: Three BUB1 and BUBR1/MAD3‐related spindle assembly checkpoint proteins are required for accurate mitosis in Arabidopsis

P31 ^comet , a member of the synaptonemal complex, participates in meiotic DSB formation in rice

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BRK1, a Bub1-Related Kinase, Is Essential for Generating Proper Tension between Homologous Kinetochores at Metaphase I of Rice Meiosis

Cited by 48 publications

References 60 publications

Simultaneous Identification of Multiple Causal Mutations in Rice

Simultaneous Identification of Multiple Causal Mutations in Rice

Retracted: Three BUB1 and BUBR1/MAD3‐related spindle assembly checkpoint proteins are required for accurate mitosis in Arabidopsis

P31 comet , a member of the synaptonemal complex, participates in meiotic DSB formation in rice

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P31 ^comet , a member of the synaptonemal complex, participates in meiotic DSB formation in rice