iProXProteomicsJikui LiuHomo Sapienshttp://www.iprox.org/page/project.html?id=IPX0002630000Liu8929@126.comPeking University Shenzhen HospitaliProX<h4>Background</h4>Long non-coding RNA colon cancer-associated transcript 1 (CCAT1) is involved in transforming multiple cancers into malignant cancer types. Previous studies underlining the mechanisms of the functions of CCAT1 primarily focused on its decoy for miRNAs (micro RNAs). However, the regulatory mechanism of CCAT1-protein interaction associated with tumor metastasis is still largely unknown. The present study aimed to identify proteome-wide CCAT1 partners and explored the CCAT1-protein interaction mediated tumor metastasis.<h4>Methods</h4>CCAT1-proteins complexes were purified and identified using RNA antisense purification coupled with the mass spectrometry (RAP-MS) method. The database for annotation, visualization, and integrated discovery and database for eukaryotic RNA binding proteins (EuRBPDB) websites were used to bioinformatic analyzing CCAT1 binding proteins. RNA pull-down and RNA immunoprecipitation were used to validate CCAT1-Vimentin interaction. Transwell assay was used to evaluate the migration and invasion abilities of HeLa cells.<h4>Results</h4>RAP-MS method worked well by culturing cells with nucleoside analog 4-thiouridine, and cross-linking was performed using 365 nm wavelength ultraviolet. There were 631 proteins identified, out of which about 60% were RNA binding proteins recorded by the EuRBPDB database. Vimentin was one of the CCAT1 binding proteins and participated in the tumor metastasis pathway. Knocked down vimetin ( VIM ) and rescued the downregulation by overexpressing CCAT1 demonstrated that CCAT1 could enhance tumor migration and invasion abilities by stabilizing Vimentin protein.<h4>Conclusion</h4>CCAT1 may bind with and stabilize Vimentin protein, thus enhancing cancer cell migration and invasion abilities.Long non-coding RNA colon cancer-associated transcript 1-Vimentin axis promoting the migration and invasion of HeLa cells.Li Zhangfu Z, Yuan Jiangbei J, Da Qingen Q, Yan Zilong Z, Qu Jianhua J, Li Dan D, Liu Xu X, Zhan Qimin Q, Liu Jikui JfalseProteome-wide identification lncRNA CCAT1 directly binding proteins by RAP-MSWe purify and identify CCAT1 binding proteins by RAP-MS. As a result, we identified out more than 600 CCAT1 binding proteins and more than 60% of them are RNA binding proteins recorded by RBPs database EuRBPDB. Among the identified CCAT1 binding proteins, Vimentin is a crucial protein. Additionally, CCAT1 could positively influence the protein level of Vimentin and enhance migration and invasion ability of cancer cells. Our data indicate that CCAT1 may bind and stabilize Vimentin enhancing the migration and invasion ability of cancer Cell.Thu Nov 26 00:00:00 GMT 2020PXD022774960637036437