Alternative splicing and differential subcellular localization of the rat FGF antisense gene product.
Alternative splicing and differential subcellular localization of the rat FGF antisense gene product.
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DOI：
10.1186/1471-2199-9-10
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发表时间：
2008-01-23
影响因子：
--
通讯作者：
Murphy PR
中科院分区：
文献类型：
--
作者：
Zhang SC;MacDonald KA;Baguma-Nibasheka M;Geldenhuys L;Casson AG;Murphy PR
GFG/NUDT是一种营养素水解酶，最初被鉴定为成纤维细胞生长因子-2反义（FGF-AS）基因的产物。虽然FGF-AS RNA被认为是FGF-2表达的反义调节剂，但编码的GFG蛋白的表达和功能在很大程度上是未知的。初级FGF-AS mRNA转录物的选择性剪接预测了包括大鼠在内的许多物种中的多种GFG亚型。在本研究中，我们专注于阐明选择性剪接大鼠GFG异构体的表达和亚细胞分布。RT-PCR和免疫组化显示，组织特异性GFG mRNA亚型的表达和GFG免疫反应的细胞质和细胞核中广泛的正常大鼠组织的亚细胞分布。FGF-2和GFG免疫反应共定位在一些，但不是所有，组织检查。计算分析确定了一个线粒体靶向序列（MTS）的N-末端的三个先前描述的rGFG亚型。共聚焦激光扫描显微镜和亚细胞分级分析表明，所有的rGFG亚型轴承MTS是专门针对线粒体，而缺乏MTS的亚型和缺失突变体定位在细胞质和细胞核。突变和缺失分析证实了预测的MTS是必要的和足够的线粒体区室化。先前的研究结果强烈支持FGF反义RNA作为FGF 2表达的调节剂的作用。目前的研究表明，反义RNA本身被翻译，并且由选择性RNA剪接产生的蛋白质异构体被分选到不同的亚细胞区室。FGF-2及其反义蛋白在许多组织中共表达，并且在某些情况下在相同细胞中共表达。跨动物物种的序列和基因组组织的强保守性表明这些转录本对的物理关联具有重要的功能意义。
GFG/NUDT is a nudix hydrolase originally identified as the product of the fibroblast growth factor-2 antisense (FGF-AS) gene. While the FGF-AS RNA has been implicated as an antisense regulator of FGF-2 expression, the expression and function of the encoded GFG protein is largely unknown. Alternative splicing of the primary FGF-AS mRNA transcript predicts multiple GFG isoforms in many species including rat. In the present study we focused on elucidating the expression and subcellular distribution of alternatively spliced rat GFG isoforms. RT-PCR and immunohistochemistry revealed tissue-specific GFG mRNA isoform expression and subcellular distribution of GFG immunoreactivity in cytoplasm and nuclei of a wide range of normal rat tissues. FGF-2 and GFG immunoreactivity were co-localized in some, but not all, tissues examined. Computational analysis identified a mitochondrial targeting sequence (MTS) in the N-terminus of three previously described rGFG isoforms. Confocal laser scanning microscopy and subcellular fractionation analysis revealed that all rGFG isoforms bearing the MTS were specifically targeted to mitochondria whereas isoforms and deletion mutants lacking the MTS were localized in the cytoplasm and nucleus. Mutation and deletion analysis confirmed that the predicted MTS was necessary and sufficient for mitochondrial compartmentalization. Previous findings strongly support a role for the FGF antisense RNA as a regulator of FGF2 expression. The present study demonstrates that the antisense RNA itself is translated, and that protein isoforms resulting form alternative RNA splicing are sorted to different subcellular compartments. FGF-2 and its antisense protein are co-expressed in many tissues and in some cases in the same cells. The strong conservation of sequence and genomic organization across animal species suggests important functional significance to the physical association of these transcript pairs.
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影响因子：
2.9
作者：
MICHAELS, ML;TCHOU, J;MILLER, JH
通讯作者：
MILLER, JH
影响因子：
64.5
作者：
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通讯作者：
KIRSCHNER, MW
影响因子：
4.1
作者：
Li, AW;Catherine, KTL;Murphy, PR
通讯作者：
Murphy, PR
影响因子：
4.1
作者：
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通讯作者：
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影响因子：
6.2
作者：
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通讯作者：
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