Identification and characterization of the lamprey Flotillin-1 gene with a role in cell adhesion

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Flotillin-1 is a kind of localize into specific cholesterol rich microdomains in cellular membranes and highly conserved lipid rafts marker protein widely distributed in animals and plants. It provides a platform for the reaction of many proteins in signal transduction, as scaffolding plays an important role in transmembrane signaling and cell adhesion. Here, Flotillin-1 protein from lamprey was identified and characterized (designated as L-Flotillin-1). After a partial cDNA sequence of L-Flotillin-1 was identified in a lamprey supraneural body cDNA library, the full-length cDNA was obtained using 3’- and 5’-rapid amplification of cDNA ends (RACE). L-Flotillin-1 encodes 424 amino acids and contains a prohibitin domain and a flotillin repetitive area. The L-Flotillin-1 protein was primarily distributed in kidney, supraneural body, gill, heart, liver and intestine via real-time PCR and immunohistochemistry assays. Immunofluorescence and western blot results showed that L-Flotillin-1 was considered to be used as a marker protein of lamprey lipid rafts and exosomes. Furthermore, overexpression of pEGFP-N1-L-Flotillin-1 induced the up-regulation of vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) mRNA levels. These results indicated that the L-Flotillin-1 gene encodes Flotillin-1 protein that was used as a conserved marker protein and may play an important role in cell adhesion, providing clues for understanding the universal functions of Flotillin-1 proteins in other species and suggesting that these proteins could serve as pattern recognition molecules in immunotherapy. We revealed that Flotillin-1 protein of lamprey overexpression in human cells plays a prevalent role in cell migration and provide new thought of treatment to diseases.HighlightsFlotillin-1 gene was identified from lamprey supraneural body cDNA library.With continuous LPS stimulation, the level of Flotillin-1 protein expression was significant increase in intestinal tissue.The overexpression of pEGFP-N1-L-Flotillin-1 induced the up-regulation of VCAM-1 and ICAM-1 mRNA levels in Hela and HepG-2 cells.

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