Screening and verification of LCMR1 interacting protein

Objective To detect and verify the interaction proteins of LCMR1 (Lung Cancer Metastasis Related Protein 1) through protein-protein interaction analysis, and provide clues for the study of LCMR1 gene function. Methods RNA of human lung cancer 95D cells was extracted as a template to construct a cDNA library. The LCMR1 plasmid and human lung cancer 95D cell cDNA library were co-transformed into yeast AH109 to screen the protein interacting with LCMR1. The interacting proteins of LCMR1 were confirmed by co-immunoprecipitation in vivo and further confirmed by GST pull-down in vitro assay. Results The 95D cell library was successfully constructed with the total RNA of human lung cancer 95D cells as a template. The LCMR1 plasmid could express the LCMR1 fusion protein correctly in yeast. Six kinds of LCMR1 interacting proteins were screened by yeast two-hybrid technique and proto-oncogene DEK was selected for validation. Co-immunoprecipitation and GST pull-down assays confirmed that DEK protein and LCMR1 protein could bind specifically in vitro and in vivo. GST pull-down assay confirmed the specific binding site of LCMR1 protein located in the N-terminal functional region of DEK. Conclusion LCMR1 interacts with DEK, and the interaction is mediated by N-terminal functional region of DEK.