SPLICING OF EFFECTOR CELL PROTEASE RECEPTOR-1 MESSENGER-RNA IS MODULATED BY AN UNUSUAL RETAINED INTRON

Effector cell protease receptor-1 (EPR-1) is a transmembrane glycoprotein receptor for factor Xa that contributes to cell surface assembly of proteolytic activities and leukocyte mitogenesis. It is now shown that membrane expression of EPR-1 is dynamically modulated by mRNA splicing. Northern hybridization analysis of EPR-1-expressing cells and genetically engineered transfectants demonstrates that this mechanism involves removal of a 451 bp intervening sequence retained in 70-90% of mature mRNA, as quantitated by polymerase chain reaction amplification and ribonuclease protection studies. Splicing of the intervening sequence occurs in a cell type-specific fashion, as judged by the constitutive membrane overexpression of EPR-1 in certain leukemic B lymphocytes and monocytic cells. Furthermore, phenotypic analysis of cell lines stably transfected with functionally spliced or unspliced EPR-1 constructs suggests a potential role of intron cis-acting sequence(s) in splicing regulation. Instead of a transmembrane receptor for factor Xa (EPR-1a), the most prevalent unspliced EPR-1 transcript generates a novel truncated protein of 110 amino acids (EPR-1b), in which a unique intron-encoded -COOH terminus carries a potential nuclear targeting signal PPQHRAKS. An antibody generated against the intron-encoded sequence of EPR-1b demonstrates prominent nuclear localization of this variant isoform in indirect immunofluorescence staining of permeabilized cells. These findings provide evidence for a novel mechanism based on high efficiency intron retention modulating factor Xa-dependent cellular effector functions.