Santangelo Philip J, Nitin Nitin, Bao Gang
Georgia Institute of Technology and Emory University, Department of Biomedical Engineering, Atlanta, Georgia 30332, USA.
J Biomed Opt. 2005 Jul-Aug;10(4):44025. doi: 10.1117/1.2011402.
The intracellular localization and specific organelle association of mRNA may reflect essential functions, stages, and stability of mRNA. We report the direct visualization of subcellular localization of K-ras and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNAs in live HDF cells using molecular beacons together with membrane-permeabilization and peptide-based delivery. Unexpectedly, we found that both K-ras and GAPDH mRNAs colocalize with mitochondria. Extensive control studies are performed, including the use of fluorescence in-situ hybridization (FISH), negative-control beacons, and the detection of colocalization of 28S ribosomal RNA with the rough endoplasmic reticulum (ER), suggesting that the mRNA localization and colocalization patterns observed in our study are true and specific. Our observation reveals intriguing subcellular associations of mRNA with organelles such as mitochondria, which may provide new insight into the transport, dynamics, and functions of mRNA and mRNA-protein interactions.
mRNA的细胞内定位和与特定细胞器的关联可能反映了mRNA的基本功能、阶段和稳定性。我们报告了使用分子信标结合膜通透化和基于肽的递送方法，在活的人皮肤成纤维细胞（HDF）中直接观察K-ras和甘油醛-3-磷酸脱氢酶（GAPDH）mRNA的亚细胞定位。出乎意料的是，我们发现K-ras和GAPDH mRNA都与线粒体共定位。我们进行了广泛的对照研究，包括使用荧光原位杂交（FISH）、阴性对照信标，以及检测28S核糖体RNA与粗面内质网（ER）的共定位，这表明我们研究中观察到的mRNA定位和共定位模式是真实且特异的。我们的观察揭示了mRNA与线粒体等细胞器之间有趣的亚细胞关联，这可能为mRNA的运输、动态变化、功能以及mRNA-蛋白质相互作用提供新的见解。
