Studies On Cytoplasmic Dynein Intermediate Chain-like Protein In Pollen Tube Of Lilium Davidii Duch

In animal and lower plant cells, cytoplasmic dynein involved in not only trafficking of subcellular organelles and target molecules, but also organizing and positioning of organelles such as Golgi apparatus. Recent studies have indicated the existence of dynein heavy chain and light chain in higher plants, but no data about dynein intermediate chain in higher plants have been reported.By SDS-PAGE and immuno-blotting, we found that a monoclonal antibody of anti-chick brain cytoplasmic dynein intermediate chain antibody could react with cytoplasmic dynein intermediate chain-like protein at 67 kDa in lily pollen.Under confocal laser scanning microscopy after immunoflurescence labeling, we found that the dynein intermediate chain-like protein appeared punctated and was co-localization partly with microtubules in cytoplasm of lily pollen tube. By SDS-PAGE and immuno-blotting, the monoclonal antibody of anti-chick brain cytoplasmic dynein intermediate chain could recognize the 67 kDa protein in purified Golgi apparatus fraction from lily pollen. Subsequently by immuno-gold labeling and transmission electron microscopy, we found that the dynein intermediate chain-like protein bound mainly to the membranes of Golgi-associated vesicles. Statistics analysis of dynein intermediate chain-like protein on Golgi-associated vesciles showed the nearly equal chance of distribution on either cis- or trans-Golgi-associated vesciles.After treatment with BFA, immunoflurescence labeling and confocal laser scanning microscopy indicated that the cytoplasmic dynein intermediate chain-like protein in lily pollen tube was insensitive to BFA treating, but spectrin-like protein was sensitive to it under the same condition.These results suggest that the cytoplasmic dynein intermediate chain-like protein exists in lily pollen, and it distributes mainly on either cis- and trans-Golgi-associated vesciles.