Translation Initiation Factor eIF3 Probably Binds with Microtubules in Mammalian Cells |
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Authors: | Shanina N. A. Ivanov P. A. Chudinova E. M. Severin F. F. Nadezhdina E. S. |
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Affiliation: | (1) Department of Molecular Biology, Moscow State University, Moscow, 119899, Russia;(2) Russian Academy of Sciences, Pushchino, Institute of Protein Research, Moscow Region, 142292, Russia;(3) Max-Planck-Institute of Molecular Cell Biology and Genetics, Dresden, 1307, Germany |
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Abstract: | Association of the translation apparatus with the cytoskeleton is essential for its transportation within the cell and probably also for translation regulation. Very little is known about the involvement of particular proteins of this association. A polypeptide homologous with the heavy chain of translation initiation factor eIF3 p170 was found earlier in a microtubule preparation from adrenal cells. Antibody A167 directed against the recombinant fragment of p170 has been generated to study eIF3 interaction with microtubules in mammalian cells. This antibody was shown to recognize a single 170-kDa polypeptide in eIF3 preparations as well as in homogenates of various cell types. A167 allowed detection of the 170-kDa polypeptide in microtubule preparation from bovine brain and confirmation of its presence in microtubule preparations from adrenal cells. As shown by immunofluorescence microscopy using A167, the 170-kDa polypeptide is mainly located in the endoplasm within numerous small and some large granules. Cell treatment with cycloheximide resulted in growth and clustering of the large granules, and partial antigen redistribution along intracellular microtubules. These new experimental data indicate that mammalian translation factor eIF3 may bind with microtubules. |
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Keywords: | translation factors eIF3 microtubules antibodies cDNA cultured cells immunofluorescence microscopy |
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