Genome wide cloning of maize meiotic recombinase Dmc1 and its functional structure through molecular phylogeny

Abstract

ABSTRACT. The development of meiotic division and associated genetic recombination paved the way for evolutionary changes. However, the secondary and tertiary structure and functional domains of many of the proteins involved in genetic recombination have not been studied in detail. We used the human Dmc1 gene product along with secondary and tertiary domain structures of Escherichia coli RecA protein to help determine the molecular structure and function of maize Dmc1, which is required for synaptonemal complex formation and cell cycle progression. The maize recombinase Dmc1 gene was cloned and characterized, using rice Dmc1 cDNA as an orthologue. The deduced amino acid sequence was used for Cloning of maize meiotic recombinase Dmc1 and its structure elaborating its 3-D structure, and functional analysis was made with the CDD software, showing significant identity of the Dmc1 gene product in Zea mays with that of Homo sapiens. Based on these results, the domains and motives of WalkerA and WalkerB as ATP binding sites, a multimer site (BRC) interface, the putative ssDNA binding L1 and L2 loops, the putative dsDNA binding helix-hairpinhelix, a polymerization motif, the subunit rotation motif, and a small N-terminal domain were proposed for maize recombinase Dmc1.

desynaptic5 carries a spontaneous semi-dominant mutation affecting Disrupted Meiotic cDNA 1 in barley

“…Although there have been a number of studies on DMC1 in other grasses, these have largely been phylogenetically or bioinformatically driven without functional studies to confirm DMC1’s role (Shimazu et al , 2001; Mikhailova et al , 2006; Devisetty et al , 2010; Etedali et al , 2011; Barakate et al , 2014). Using super-resolution immunocytology we show that des5 in barley has a deleterious effect on synapsis and crossing over, and that we can recapitulate the meiotic and semi-sterile mutant phenotypes using RNAi knockdowns in transgenic plants.…”

Simultaneous TALEN-mediated knockout of chrysanthemum DMC1 genes confers male and female sterility

“…1, Supplementary Table S2). This target sequence includes the multimer site (BRC) interface located upstream of the Walker B motif 20 . production of CmDMC1 knockout plants using TALEN-mediated targeted mutagenesis.…”

“…1 , Supplementary Table S2 ). This target sequence includes the multimer site (BRC) interface located upstream of the Walker B motif 20 .

Figure 1 Partial genomic DNA sequences of the six CmDMC1 genes of Chrysanthemum morifolium cultivar ‘Shuho-no-chikara’.

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“…They used transgenic chrysanthemum lines harboring more than four or five copies of a gene for yellow-green fluorescent protein from Chiridius poppei ( CpYGFP ) as materials for gene disruption, and edited all the CpYGFP -transgene alleles simultaneously. In this study, we isolated six CmDMC1 loci and intended to mutagenize the BRC multimer interface just upstream of the Walker B motif 20 , since Walker A and/or B motifs are functionally essential ATP-binding sites 40 . Disruption of these motifs by frameshift mutations could destroy DMC1 function, resulting in abnormal meiosis and, consequently, a male and female sterility phenotype.…”

“…In contrast, the allele harboring a deletion of three nucleotides potentially generates deletion of a single amino acid in the respective DMC1 proteins encoded by a single allele of CmDMC1d (lines SHa#13 and SHf#20) or CmDMC1e (SHb#14 and YSc#27). In these four lines, each CmDMC1d or CmDMC1e mutant protein carries a deletion of Lys at position 215 (K215) located in the BRC multimer interface domain, which was proposed to be involved in monomer–monomer interaction 20 . This Lys residue—in equivalent positions in DMC1 proteins—is shared among mammals and higher plants 49 that bear both BRCA and DMC1 genes.…”