Intron-mediated expression of the human neuropeptide Y Y1 receptor.
The Neuropeptide Y (NPY) family of neuropeptides exert their function through a family of heptahelical G-protein coupled receptors regulating essential physiological processes. A 97 base pair intron (intron IV) intervenes the coding sequence of the human NPY Y1 receptor (hY1) gene and was found frequently retained at variable levels in poly A+ mRNA isolated from multiple human tissues. When included in hY1 expression vectors, either in its natural position or 5' of the hY1 cDNA, intron IV mediated a significant increase of both hY1 mRNA and corresponding functional receptor protein in transfected mammalian cells, implying an in vivo regulatory function of the endogenous intron. Our results further indicate that the nuclear history of the hY1 pre-mRNA influence ectopic hY1 production through post-transcriptional mechanisms and argues against intron IV acting as a transcriptional enhancer as well as the possibility that a putative hY1 related 5TM accessory protein encoded by the non-spliced hY1 mRNA would facilitate hY1 production on a post-translational level.
Marklund
Ulrica
U
Department of Molecular Biology, AstraZeneca R&D, Tvistevägen 48, S-907 36 Umeå, Sweden.
Byström
Mona
M
Gedda
Karin
K
Larefalk
Asa
A
Juneblad
Kristina
K
Nyström
Susanne
S
Ekstrand
A Jonas
AJ
D020121:5' Untranslated Regions; D000818:Animals; D001667:Binding, Competitive; D015152:Blotting, Northern; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000737:chemistry; D017353:Gene Deletion; D015870:Gene Expression / Q000187:drug effects; D049452:Green Fluorescent Proteins; D006801:Humans; D007438:Introns / Q000235:genetics*; D008164:Luminescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D009479:Neuropeptides / Q000737:chemistry / Q000494:pharmacology; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012326:RNA Splicing / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D017476:Receptors, Neuropeptide Y / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D014162:Transfection
D020121:5' Untranslated Regions; D017931:DNA Primers; D008164:Luminescent Proteins; D009479:Neuropeptides; D010446:Peptide Fragments; D012333:RNA, Messenger; D017476:Receptors, Neuropeptide Y; C097212:neuropeptide Y-Y1 receptor; D049452:Green Fluorescent Proteins; D000242:Cyclic AMP; D019204:GTP-Binding Proteins