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Publication : Molecular cloning and subcellular localization of three GTP-binding proteins of the rab subfamily.

First Author  Olkkonen VM Year  1993
Journal  J Cell Sci Volume  106 ( Pt 4)
Pages  1249-61 PubMed ID  8126105
Mgi Jnum  J:24141 Mgi Id  MGI:71898
Doi  10.1242/jcs.106.4.1249 Citation  Olkkonen VM, et al. (1993) Molecular cloning and subcellular localization of three GTP-binding proteins of the rab subfamily. J Cell Sci 106(Pt 4):1249-61
abstractText  Small GTPases of the rab subfamily are involved in regulation of intracellular membrane transport events. We recently used a PCR approach to isolate short cDNA fragments of a number of novel rab sequences. These PCR fragments have not been used with cDNA library screening and PCR-based techniques to clone the cDNAs encoding three of these proteins, rab12, rab22, and rab24. By northern blot analysis, the messages were found to be present in a wide variety of mouse tissues. However, quantitative differences in the mRNA levels between the tissues were detected. We determined the subcellular localization of the GTPases by expressing the c-myc epitope-tagged proteins with the Semliki Forest virus and the vaccinia T7 vector systems. Transiently expressed rab12 was localized to the Golgi complex. This localization was confirmed using a polyclonal anti-peptide antibody detecting the endogenous protein in BHK cells. rab22 expressed from the cDNA was localized to endosomal compartments and to the plasma membrane. After longer periods of expression, the protein was found on abnormally large perinuclear endosomal structures, suggesting that it is a potent regulator of events in the endocytic pathway. Finally, rab24 was found in the endoplasmic reticulum/cis-Golgi region and on late endosomal structures. The localization of rab24 may indicate its involvement in autophagy-related processes.
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