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Post-Transcriptional Maturation of the S Receptor Kinase of Brassica Correlates with Co-Expression of the S-Locus Glycoprotein in the Stigmas of Two Brassica Strains and in Transgenic Tobacco Plants

机译:芸苔属植物S受体激酶的转录后成熟与两个芸苔属植物的柱头和转基因烟草植物中S-基因座糖蛋白的共表达有关

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摘要

The S-locus-encoded S receptor kinase (SRK) is an intrinsic plasma membrane protein that is viewed as the primary stigma determinant of specificity in the self-incompatibility response of Brassica spp. We analyzed two self-compatible mutant strains that express low levels of the S-locus glycoprotein (SLG), a cell wall-localized protein also encoded at the S locus that is coordinately expressed with SRK. We found that mutant stigmas synthesized wild-type levels of SRK transcripts but failed to produce SRK protein at any of the developmental stages analyzed. Furthermore, SRK was shown to form aberrant high-molecular mass aggregates when expressed alone in transgenic tobacco (Nicotiana tabacum) plants. This aggregation was prevented in tobacco plants that co-expressed SRK and SLG, but not in tobacco plants that co-expressed SRK and SLR1, an SLG-related secreted protein not encoded at the S locus. In analyses of protein extracts under reducing and non-reducing conditions, evidence of intermolecular association was obtained only for SLG, a fraction of which formed disulfide-linked oligomers and was membrane associated. The data indicate that, at least in plants carrying the S haplotypes we analyzed, SRK is an inherently unstable protein and that SLG facilitates its accumulation to physiologically relevant levels in Brassica stigmas.
机译:S-位点编码的S受体激酶(SRK)是一种固有的质膜蛋白,被视为甘蓝属植物自交不亲和反应中特异性的主要柱头决定因素。我们分析了两个自我兼容的突变株,它们表达低水平的S-基因座糖蛋白(SLG),一种在S基因座上也与SRK协同表达的细胞壁定位蛋白。我们发现突变的柱头合成了SRK转录本的野生型水平,但在所分析的任何发育阶段均未产生SRK蛋白。此外,当SRK在转基因烟草(Nicotiana tabacum)植物中单独表达时,它显示出异常的高分子物质聚集体。在共表达SRK和SLG的烟草植物中防止了这种聚集,但在共表达SRK和SLR1的烟草植物中则阻止了这种聚集,SRK和SLR1是S位点未编码的SLG相关分泌蛋白。在还原和非还原条件下对蛋白质提取物进行分析时,仅SLG获得了分子间缔合的证据,SLG中的一部分形成了二硫键连接的低聚物,并且与膜相关。数据表明,至少在带有我们分析的S单倍型的植物中,SRK是一种固有的不稳定蛋白,SLG促进其在芸苔属柱头上的积累达到生理相关水平。

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