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Characterization and intracellular localization of the Rok1 protein involved in yeast cell division
1Department of Microbiology, College of Natural Sciences, Chungnam National University, Taejeon, Korea.
Abstract:
The ROK1 gene is essential for the cell cycle progression in Saccharomyces cerevisiae. ROK1 has been predicted to encode an ATP-dependent RNA helicase of the DEAD-box family. We have analyzed the ROK1 gene expression both at the protein and RNA levels. Polyclonal antibodies were raised against trpE::rok1 hybrid proteins and were affinity purified by using lacZ::rok1 hybrid proteins. Western blot experiments using anti-Rok1 antibodies revealed a single protein band of 64 kDa which is an expected size from the Rok1 amino acid sequence. Indirect immuno-fluorescence showed that the Rok1 protein is localized predominantly to the cytoplasm of the vegetatively growing cells. We have detected immunocross-reactive homologs of Rok1p in Candida albicans and Drosophila melanogaster.