Biopolymers and cell. 2010; 26 (6): 512 - 516
Novel antibodies against RCD-8 as a tool to study processing bodies
D. O. Gudkova1,2, G. G. Panasyuk1, I. O. Nemazanyy1, V. V. Filonenko1
1Institute of Molecular Biology and Genetics NAS of Ukraine, 150, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
2Taras Shevchenko Kyiv National University, 64, Volodymyrska Str., Kyiv, Ukraine, 01033
Aim. To develop the model system for processing bodies (PBs) state monitoring and accomplish it in the future as a possible read-out of mTOR activity in mammalian cells. Methods. In course of this study we raised polyclonal antibodies against one of the PBs scaffold proteins – RCD-8 and employed cell imaging technique. Results. It has been shown that the obtained antibodies recognize the intracellular structures, namely PBs. The detected protein co-localized with known marker of PBs – DCP1a, and partly with marker of SGs – CPEB. Conclusions. Based on changes of PBs number and size in cells after exposure to known inductors or inhibitors of PB formation we prove the specificity of generated antibodies and possibility of their application for studies on the processing bodies dynamics controlled by mTOR-dependent signaling.
Key words: mRNA processing, decapping, processing bodies, immunocytochemistry.