Biopolym. Cell. 2010; 26(6):512-516.
Short Communications
Novel antibodies against RCD-8 as a tool to study processing bodies
1, 2Gudkova D. O., 1Panasyuk G. G., 1Nemazanyy I. O., 1Filonenko V. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Taras Shevchenko National University of Kyiv
    64, Volodymyrska Str., Kyiv, Ukraine, 01601


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.
Keywords: mRNA processing, decapping, processing bodies, immunocytochemistry


[1] Coller J., Parker R. Eukaryotic mRNA decapping Annu. Rev. Biochem 2004 73:861–890.
[2] Cougot N., van Dijk E., Babajko S., Seraphin B. «Cap-tabolism» Trends Biochem. Sci 2004 29, N 8:436–444.
[3] Buchan J.R, Parker R. Eukaryotic stress granules: the ins and outs of translation Mol. Cell 2009 36, N 6:932–941.
[4] Wilusz C., Wormington M., Peltz S. The cap-to-tail guide to mRNA turnover Nat. Revs Mol. Cell Biol 2001 2, N 4:237–246.
[5] Averous J., Proud C. G. When translation meets transformation: the mTOR story Oncogene 2006 25, N 48:6423– 6435.
[6] Liu L., Li F., Cardelli J. A., Martin K. A., Blenis J., Huang S. Rapamycin inhibits cell motility by suppression of mTORmediated S6K1 and 4E-BP pathways Oncogene 2006 25, N 53:7029–7070.
[7] Sun S., Rosenberg L. M., Wang X., Zhou Z., Yue P., Fu H., Khuri F. R. Activation of akt and eIF4E survival pathways by rapamycin-mediated mammalian target of rapamycin inhibition Cancer Res 2005 65, N 16:7052–7058.
[8] Stinton L. M., Eystathioy T., Selak S., Chan E. K., Fritzler M. J. Autoantibodies to protein transport and messenger RNA processing pathways: endosomes, lysosomes, Golgi complex, proteasomes, assemblyosomes, exosomes, and GW bodies Clin. Immunol 2004 110, N 1:30–44.
[9] Parker R., Sheth U. P bodies and the control of mRNA translation and degradation Mol. Cell 2007 25, N 5:635–646.
[10] Garcia-Lozano J. R., Gonzales-Escribano M. F., Wichmann I., Nunez-Roldan A. Cytoplasmic detection of a novel protein containing a nuclear localization sequence by human autoantibodies Clin. Exp. Immunol 1997 107, N 3:501–506.
[11] Yu J. H., Yang W.-H., Gulich T., Bloch K. D., Bloch D. Ge-1 is a central component of the mammalian cytoplasmic mRNA processing body RNA 2005 11, N 12:1795–1802.
[12] Bloch D. B., Gulick T., Bloch K. D., Yang W.-H. Processing body autoantibodies reconsidered RNA 2006 12, N 5:707–709.
[13] Fenger-Gron M., Fillman C., Norrild B., Lykke-Andersen J. Multiple processing body factors and the ARE binding protein TTP activate mRNA decapping Mol. Cell 2005 20, N 6:905–915.
[14] Nemazanyy I., Breus O., Gout I., Filonenko V., Panasyuk G. Generation and characterization of monoclonal antibodies to mTOR kinase Hybridoma 2008 27, N 5:395–399.
[15] Wilczynska A., Aigueperse C., Kress M., Dautry F., Weil D. The translational regulator CPEB1 provides a link between dcp1 bodies and stress granules J. Cell Sci 2005 118, N 5:981–992.
[16] Mollet S., Cougot N., Wilczynska A., Dautry F., Kress M., Bertrand E., Weil D. Translationally repressed mRNA transiently cycles through stress granules during stress Mol. Biol. Cell 2008 19, N 10:4469–4479.
[17] Cavigelli M., Li W., Lin A., Su B., Yoshioka K., Karin M. The tumor promoter arsenite stimulates AP-1 activity by inhibiting a JNK phosphatase The EMBO J 1996 15, N 22:6269–6279.