title: Characterization of a novel splicing variant in the RAPTOR gene.
abstract: The mammalian target of rapamycin (mTOR) plays an essential role in the regulation of cell growth, proliferation and apoptosis. Raptor, the regulatory associated protein of mTOR, is an important member in this signaling pathway. In the present report,we identified and characterized a novel splicing variant of this gene, RAPTOR v2, in which exons 14-17, 474 bp in total, are omitted from the mRNA. This deletion does not change the open reading frame, but causes a nearly complete absence of HEAT repeats, which were shown to be involved in the binding of mTOR substrates. Real time PCR performed on 48 different human tissues demonstrated the ubiquitous presence of this splice variant. Quantification of mRNA levels in lymphoblastoid cell lines (LCL) from 56 unrelated HapMap individuals revealed that the expression of this splicing form is quite variable. One synonymous SNP, rs2289759 in exon 14, was predicted by ESEfinder to cause a significant gain/loss of SRp55 and/or SF2/ASF binding sites, and thus potentially influence splicing. This prediction was confirmed by linear regression analysis between the ratio of RAPTOR v2 to total RAPTOR mRNA levels and the SNP genotype in the above 56 individuals (r=0.281 and P=0.036). Moreover, the functional evaluation indicated that this splicing isoform is expected to retain the ability to bind mTOR, but is unlikely to bind mTOR substrates, hence affecting signal transduction and further cell proliferation.
authors.affiliation: Department of Human Genetics, University of Chicago, Chicago, IL 60637, USA. csun1@bsd.uchicago.edu
authors.forename: Chang
authors.initials: C
authors.lastname: Sun
authors.forename: Catherine
authors.initials: C
authors.lastname: Southard
authors.forename: Anna
authors.initials: A
authors.lastname: Di Rienzo
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D017398:Alternative Splicing / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000076223:Regulatory-Associated Protein of mTOR
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D011506:Proteins; D012333:RNA, Messenger; C463659:RPTOR protein, human; D000076223:Regulatory-Associated Protein of mTOR