Petersen Nikolaj H T, Joensen Jan, McKinney Lea V, Brodersen Peter, Petersen Morten, Hofius Daniel, Mundy John
Department of Biology, Copenhagen Biocenter, Copenhagen N, Denmark.
J Plant Physiol. 2009 Apr 1;166(6):661-6. doi: 10.1016/j.jplph.2008.08.003. Epub 2008 Oct 8.
The Arabidopsis ACD11 gene encodes a sphingosine transfer protein and was identified by the accelerated cell death phenotype of the loss of function acd11 mutant, which exhibits heightened expression of genes involved in the disease resistance hypersensitive response (HR). We used ACD11 as bait in a yeast two-hybrid screen of an Arabidopsis cDNA library to identify ACD11 interacting proteins. One interactor identified is a protein of unknown function with an RNA recognition motif (RRM) designated BPA1 (binding partner of ACD11). Co-immunoprecipitation experiments confirmed the ACD11-BPA1 interactions in vivo and in vitro. Two other ACD11 interactors (PRA7 and PRA8) are homologous to each other and to mammalian PRA1, and both were subsequently shown to interact with BPA1 in yeast. A fourth interactor (VAP27-1) is homologous to mammalian VAP-A, and was found to interact more strongly with a homolog of ACD11 than ACD11 itself. All interactors were shown to be associated with membrane fractions, suggesting that ACD11 function could be related to the regulation of membrane compartments.
拟南芥ACD11基因编码一种鞘氨醇转移蛋白，通过功能缺失型acd11突变体的加速细胞死亡表型得以鉴定，该突变体表现出参与抗病超敏反应（HR）的基因表达增强。我们以ACD11为诱饵，在拟南芥cDNA文库的酵母双杂交筛选中鉴定与ACD11相互作用的蛋白。鉴定出的一个相互作用蛋白是一种功能未知的蛋白，带有RNA识别基序（RRM），命名为BPA1（ACD11的结合伴侣）。免疫共沉淀实验证实了ACD11与BPA1在体内和体外的相互作用。另外两个ACD11相互作用蛋白（PRA7和PRA8）彼此同源且与哺乳动物PRA1同源，随后二者均被证明在酵母中与BPA1相互作用。第四个相互作用蛋白（VAP27-1）与哺乳动物VAP-A同源，并且发现它与ACD11的一个同源物的相互作用比与ACD11本身的相互作用更强。所有相互作用蛋白均显示与膜组分相关，这表明ACD11的功能可能与膜区室的调节有关。
