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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Localization of mRNAs coding for peroxisomal proteins in the yeast, Saccharomyces cerevisiae
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Localization of mRNAs coding for peroxisomal proteins in the yeast, Saccharomyces cerevisiae

机译：编码酿酒酵母中过氧化物酶体蛋白的mRNA的定位

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摘要

Targeted mRNA trafficking and local translation may play a significant role in controlling protein localization. Here we examined for the first time the localization of all (≈50) mRNAs encoding peroxisomal proteins (mPPs) involved in peroxisome biogenesis and function. By using the bacteriophage MS2-CP RNA-binding protein (RBP) fused to multiple copies of GFP, we demonstrated that ＞40 endogenously expressed mPPs tagged with the MS2 aptamer form fluorescent RNA granules in vivo. The use of different RFP-tagged organellar markers revealed 3 basic patterns of mPP granule localization: to peroxisomes, to the endoplasmic reticulum (ER), and nonperoxisomal. Twelve mPPs (i.e., PEX1, PEX5, PEX8, PEX11-15, DCI1. NPY1. PCS60, and POX1) had a high percentage (52%-80%) of mRNA colocalization with peroxisomes. Thirteen mPPs (i.e., AAT2, PEX6. MDH3, PEX28, etc.) showed a low percentage (30%-42%) of colocalization, and 1 mPP (PEX3) preferentially localized to the ER. The mPPs of the nonperoxisomal pattern (i.e., GPD1, PCD1, PEX7) showed 30% colocalization. mPP association with the peroxisome or ER was verified using cell fractionation and RT-PCR analysis. A model mPP, PEX14 mRNA, was found to be in close association with peroxisomes throughout the cell cycle, with its localization depending in part on the 3-UTR, initiation of translation, and the Puf5 RBP. The different patterns of mPP localization observed suggest that multiple mechanisms involved in mRNA localization and translation may play roles in the importation of protein into peroxisomes.
机译：靶向的mRNA转运和局部翻译可能在控制蛋白质定位中起重要作用。在这里，我们首次检查了所有编码过氧化物酶体生物发生和功能的过氧化物酶体蛋白（mPPs）的所有（≈50）mRNA的定位。通过使用噬菌体MS2-CP RNA结合蛋白（RBP）与GFP的多个副本融合，我们证明了带有MS2适体标记的＞ 40个内源表达的mPP在体内形成了荧光RNA颗粒。使用不同的RFP标记的细胞器标记物揭示了mPP颗粒定位的3种基本模式：过氧化物酶体，内质网（ER）和非过氧化物酶体。十二个mPP（即PEX1，PEX5，PEX8，PEX11-15，DCI1，NPY1，PCS60和POX1）具有与过氧化物酶体共定位的mRNA高比例（52％-80％）。十三个mPP（即AAT2，PEX6，MDH3，PEX28等）显示出较低的共定位百分比（30％-42％），并且有1个mPP（PEX3）优先定位于ER。非过氧化物酶体模式的mPPs（即GPD1，PCD1，PEX7）显示 30％的共定位。使用细胞分级分离和RT-PCR分析验证了mPP与过氧化物酶体或ER的关联。已发现模型mPP PEX14 mRNA在整个细胞周期中与过氧化物酶体密切相关，其定位部分取决于3-UTR，翻译起始和Puf5 RBP。观察到的mPP定位的不同模式表明，参与mRNA定位和翻译的多种机制可能在蛋白质导入过氧化物酶体中起作用。

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