非受体酪氨酸激酶c-abl调节微管组装的机理研究-mechanism of non-receptor tyrosine kinase c - abl regulating microtubule assembly.docxVIP

signals. PLK1 is a member of Polo-like kinases which is widely expressed in eukaryotic cells. As a serine/threonine kinase, PLK1 plays important roles in cell cycle, cell proliferation, tumorigenesis and other aspects. They are both involved in the formation of centrosomes and spindles. FOXM1 which is one of the substrates of PLK1 kinase, can regulate the express of PLK1. They regulate series of cell cycle proteins and factors by interactions to influence cell cycle.In this study, we demonstrated that c-Abl interacted with PLK1 and regulated its phosphorylation through immunobloting and co-immunoprecipiation, so PLK1 is a substrate of c-Abl. We synchronized cells and found that the interaction between c-Abl and PLK1 and the phosphorylation of PLK1 changed with the cell cycle. We demonstrated that c-Abl could promote the transcription of PLK1 through Fluorescence Quantitative PCR and Report Gene Detection technology; c-Abl could inhibit the degradation of FOXM1 through Pulse-chase; c-Abl could enhance the protein level of FOXM1 which was dependent on its activity through Westernblot. Thus c-Abl kinase influenced the expression and protein stability of FOXM1 indirectly by interacting with and phosphorylation of one of its substrate-PLK1.In this study, we separated the polymerized and soluble tubulins, and tested their protein levels by immunobloting. We found that the proportion of soluble ones increased and the polymerized ones decreased after c-Abl was knockdown. Then we found that the polymerization extent was decreased sharply when c-Abl was knockdown after separated the microtubules in physiological state, which showed that c-Abl could inhibit the depolymerization of microtubules. Through immuno-fluorescent assay, we found that the microtubule structure was dispersed when c-Abl was knockdown, which showed the assembly of microtubules was influenced by c-Abl. Besides, we compared the tubulin levels between wild type cell line and the c-Abl-knockdown one. We found