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Dual Targeting to Mitochondria and Plastids of AtBT1 and ZmBT1, Two Members of the Mitochondrial Carrier Family

机译:双重靶向线粒体载体家族的两个成员AtBT1和ZmBT1的线粒体和质体

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摘要

Zea mays and Arabidopsis thaliana Brittle 1 (ZmBT1 and AtBT1, respectively) are members of the mitochondrial carrier family. Although they are presumed to be exclusively localized in the envelope membranes of plastids, confocal fluorescence microscopy analyses of potato, Arabidopsis and maize plants stably expressing green fluorescent protein (GFP) fusions of ZmBT1 and AtBT1 revealed that the two proteins have dual localization to plastids and mitochondria. The patterns of GFP fluorescence distribution observed in plants stably expressing GFP fusions of ZmBT1 and AtBT1 N-terminal extensions were fully congruent with that of plants expressing a plastidial marker fused to GFP. Furthermore, the patterns of GFP fluorescence distribution and motility observed in plants expressing the mature proteins fused to GFP were identical to those observed in plants expressing a mitochondrial marker fused to GFP. Electron microscopic immunocytochemical analyses of maize endosperms using anti-ZmBT1 antibodies further confirmed that ZmBT1 occurs in both plastids and mitochondria. The overall data showed that (i) ZmBT1 and AtBT1 are dually targeted to mitochondria and plastids; (ii) AtBT1 and ZmBT1 N-terminal extensions comprise targeting sequences exclusively recognized by the plastidial compartment; and (iii) targeting sequences to mitochondria are localized within the mature part of the BT1 proteins.
机译:玉米和拟南芥脆性1(分别为ZmBT1和AtBT1)是线粒体载体家族的成员。尽管推测它们仅局限在质体的包膜中,但对稳定表达ZmBT1和AtBT1的绿色荧光蛋白(GFP)融合体的马铃薯,拟南芥和玉米植物进行共聚焦荧光显微镜分析表明,这两种蛋白对质体和蛋白质具有双重定位。线粒体。在稳定表达ZmBT1和AtBT1 N端延伸的GFP融合体的植物中观察到的GFP荧光分布模式与表达与GFP融合的质体标记的植物完全一致。此外,在表达与GFP融合的成熟蛋白的植物中观察到的GFP荧光分布和运动的模式与在表达与GFP融合的线粒体标记的植物中观察到的模式相同。使用抗ZmBT1抗体对玉米胚乳进行电子显微镜免疫细胞化学分析,进一步证实ZmBT1既存在于质体中也存在于线粒体中。总体数据显示:(i)ZmBT1和AtBT1双重靶向线粒体和质体; (ii)AtBT1和ZmBT1 N端延伸包含仅由质体区隔识别的靶向序列; (iii)针对线粒体的靶向序列位于BT1蛋白的成熟部分内。

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