Research On The Interaction Between MARVELD1 And Nucleolin & Their Impact In DNA Damage

Oncogene, tumor-suppressor gene and other related genes have a very important role in the cancer occurrence and development. The MARVELD1 is a novel functional gene which is found by our laboratory when exploreing carcinogenesis. It is so called MARVELD1 because that it has the MARVEL(MAL and related proteins for vesicle trafficking and membrane link) domain. Our reseach has shown that MARVELD1 is a tumor-suppressor gene, which can causes the inhibition of tumor proliferation and lead to cell cycle arrest. Moreover, MARVELD1 can interact with a variety of proteins.According to the previous experimental results, MARVELD1 has protein-protein interactions with Nucleolin, which is the highest expressed protein in the nucleus. They are all most located in the nucleus, participating in the DNA damage repair process. So we use DNA damage induced technique, choosing multiple methods, to study the interaction between MARVELD1 and nucleolin.First, we analyzed the colocalization of MARVELD1 and Nucleolin in He La cells by LSCM. Then we used Co-Immunoprecipitation assay and GST-pulldown assay to study the interaction of MARVELD1 with Nucleolin. To confirm their interacting domains, we constructed the GST-nucleolin recombinant plasmids. We detected the change of MARVELD1 and Nucleolin expression after inducing DNA damage by WB. Using the technique of subcellular components separation and RNA interference, we studied the effect of MARVELD1 on the subcellular localization of Nucleolin.Results showd that MARVELD1 and Nucleolin is obviously co-localized after UV irradiation. They have protein-protein interaction, which is dynamically changing after DNA damage. The expression of nucleolin does not happen in conformity with MARVELD1 on protein level. The expression of nucleoin begin to increase when the expreesion of MARVELD1 is though declining. Subcellular components separation and RNA interference showed that MARVELD1 has effect on the subcellular localization of Nucleolin.In conclusion, we confirmed the protein-protein interaction of MARVELD1 with Nucleolin and their interacting domain, and they are all involved with the DNA damage response process, which is probably maintained by MARVELD1 affecting the subcellular localization of Nucleolin.