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After digestion, zinc (Zn) was not detected (ND) in the GST sample, whereas the GST–CRC sample did contain zinc. The input for proteinase K digestion of GST and GST–CRC, respectively, is shown. (D) Zinc content of GST and GST–CRC proteins determined with zincon dye. Br, bromine Ca, calcium Cr, chromium Zn, zinc Cu, copper Pb, lead Sn, tin. The values were obtained by inductively coupled plasma mass spectrometry analysis and are shown in μg metal g −1 solution. (C) Illustration showing the metal content of a GST–CRC fusion protein solution (black columns) and in the buffer blank (white columns).
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(B) Schematic of the cysteine-rich motif of the D. (A) Schematic of the CRC domain of TCX proteins. Zinc binding by the CRC domain and structures of the conserved cysteine-rich domains in TCX and E(z) type proteins. Despite the similarity between TCX and E(z) type proteins, a clear distinction can be made since E(z) proteins lack the RNPXAFXPK motif ( Fig. These proteins belong to the polycomb group of proteins, which are involved in maintaining stable repression of gene expression through cell divisions and have functions in the regulation of cell proliferation ( Reyes and Grossniklaus, 2003). thaliana these include CURLY LEAF ( Goodrich et al., 1997 Kim et al., 1998) and MEDEA ( Grossniklaus et al., 1998 Kiyosue et al., 1999). As previously noted ( Cvitanich et al., 2000 Hauser et al., 2000 Song et al., 2000), the cysteine-rich domains most similar to the CRC domain are found in the Drosophila melanogaster Enhancer of zeste protein and its homologues. Here, the term CRC domain will be used, and proteins containing the CRC domain will be called TCX proteins. This domain has previously been named the CRC domain, the TCR motif, and the CHC domain ( Cvitanich et al., 2000 Song et al., 2000 Sutou et al., 2003). Both Hstesmin and AtTSO1 localize to the nucleus and contain two cysteine-rich CXC motifs (pfam03638) with the consensus sequence CXCX 4CX 3YCXCX 6CX 3CXCX 2C separated by a region of variable length containing the short conserved sequence RNPXAFXPK ( Fig. ‘Tso’ means ‘ugly’ in Chinese and refers to the appearance of tso1 mutant flowers. Concomitantly, mutant alleles of the Arabidopsis thaliana TSO1 gene causing defects in flower and ovule development were identified ( Hauser et al., 2000 Song et al., 2000). The human TESMIN gene was originally identified by its specific expression in testes, but subsequently it was also detected at specific stages of ovary development ( Sugihara et al., 1999 Sutou et al., 2003 Olesen et al., 2004). Male germ cells are known as sperm cells, and are produced in the testes of animals and anthers of plants, respectively. In flowering plants, the ovary is embedded in the carpel and contains ovules, which develop into seeds after fertilization. Female germ cells are produced in the ovary. During this process haploid germ cells have to be produced, and the development of specialized organs, which facilitate germ cell production, is a common challenge to plants and animals. Most animals and flowering plants are capable of sexual reproduction. Meristem, ovule, pollen, retinoblastoma, SynMuv Introduction This was confirmed by analysis of novel tso1 T-DNA insertion alleles where the development of both pollen and ovules was affected. Their expression ratios indicated that pollen, in addition to ovules, would be sensitive to loss of TSO1 function. In addition, the phylogenetic and expression analysis of the TCX genes suggested an overlap in function between AtTSO1 and the related gene AtTCX2.
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It is shown that the CRC domain binds zinc, offering an explanation for the severity of tso1 alleles where cysteine residues are affected. In the present study, the tesmin/TSO1-like CXC (TCX) proteins are characterized at the biochemical, genomic, transcriptomic, and functional level to address this question. The tso1 mutants described so far carry point mutations in a conserved cysteine-rich domain, the CRC domain, but the reason for the range of phenotypes observed is poorly understood. Mild tso1 mutant alleles influence only ovule development, whereas strong alleles have an effect on all floral tissues and cause cell division defects. The Arabidopsis thaliana TSO1 gene is involved in both these processes. Development of reproductive tissue and control of cell division are common challenges to all sexually reproducing eukaryotes.