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玉米MuDR转座子编码的MURA和MURB蛋白的亚细胞定位

Subcellular localization of MURA and MURB proteins encoded by the maize MuDR transposon.

作者信息

Ono Akemi, Kim Soo-Hwan, Walbot Virginia

机构信息

Department of Biological Sciences, Stanford University, CA 94305-5020, USA

出版信息

Plant Mol Biol. 2002 Nov;50(4-5):599-611. doi: 10.1023/a:1019970206057.

Abstract

MuDR controls transposition of the Mu transposable element family in Zea mays L. It produces two major transcripts: mudrA and mudrB, mudrA encodes the MURA transposase, but no specific function has been ascribed to mudrB, which lacks strong homology to known genes. Using transient expression assays in onion epidermal cells, we defined three monopartite nuclear localization signals (NLSs) of MURA; each was functionally sufficient for nuclear targeting of MURA:GUS fusion proteins. Interestingly, one NLS (NLS-A3) is produced by the splicing of the third intron. In contrast, there were no clear NLS in MURB, and the major form of MURB aggregated in the cytoplasm. Self-interaction of MURA and of MURB was also shown in a yeast two-hybrid assay. To test whether interactions of MURA and MURB can occur at the level of protein translocation into the nucleus, a cytoplasmically localized MURB:GFP was co-expressed with MURA or with the GUS fusion proteins. Co-expression did not change the localization pattern of either MURA or MURB; MURA and MURB do not detectably interact in a yeast two-hybrid assay. These results suggest that MURA and MURB do not mutually affect their localization, at least in the forms examined here.

摘要

MuDR控制玉米中Mu转座元件家族的转座。它产生两种主要转录本:mudrA和mudrB,mudrA编码MURA转座酶,但mudrB尚未明确其特定功能,它与已知基因缺乏强同源性。通过在洋葱表皮细胞中进行瞬时表达分析,我们确定了MURA的三个单部分核定位信号(NLS);每个信号对于MURA:GUS融合蛋白的核靶向功能都足够。有趣的是,一个NLS(NLS-A3)是由第三个内含子的剪接产生的。相比之下,MURB中没有明显的NLS,并且MURB的主要形式聚集在细胞质中。酵母双杂交试验也显示了MURA和MURB的自我相互作用。为了测试MURA和MURB的相互作用是否能在蛋白质转运到细胞核的水平上发生,将细胞质定位的MURB:GFP与MURA或GUS融合蛋白共表达。共表达并没有改变MURA或MURB的定位模式;在酵母双杂交试验中,MURA和MURB没有可检测到的相互作用。这些结果表明,至少在这里所检测的形式中,MURA和MURB不会相互影响它们的定位。

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