title: Maintenance of Pdx1 mRNA translation in islet β-cells during the unfolded protein response.
abstract: In type 1 diabetes, proinflammatory cytokines secreted by infiltrating immune cells activate the unfolded protein response (UPR) in islet β-cells, which leads to attenuation of global mRNA translation. Under such conditions, privileged mRNAs required for adaptation to the prevailing stress are maintained in an actively translated state. Pdx1 is a β-cell transcription factor that is required for the adaptive UPR, but it is not known how translation of its mRNA is maintained under these conditions. To study translation, we established conditions in vitro with MIN6 cells and mouse islets and a mixture of proinflammatory cytokines (IL-1β, TNF-α, and IFN-γ) that mimicked the UPR conditions seen in type 1 diabetes. Cell extracts were then subjected to polyribosome profiling to monitor changes to mRNA occupancy by ribosomes. Similar to other privileged mRNAs (Atf4 and Chop), Pdx1 mRNA remained partitioned in actively translating polyribosomes under the UPR, whereas the mRNA encoding a proinsulin-processing enzyme (Cpe) and others partitioned into inactively translating monoribosomes. Bicistronic luciferase reporter analyses revealed that the distal portion of the 5'-untranslated region of mouse Pdx1 (between bp -105 to -280) contained elements that promoted translation under both normal and UPR conditions, and this region exhibited conserved sequences and secondary structure similar to those of other known internal ribosome entry sites. Our findings suggest that Pdx1 protein levels are maintained in the setting of the UPR, in part, through elements in the 5'-untranslated region that confer privileged mRNA translation in a 5'-7-methylguanylate cap-independent manner.
authors.affiliation: Department of Cellular and Integrative Physiology (A.T.T., R.G.M.), Department of Pediatrics and the Herman B Wells Center for Pediatric Research (B.M., S.A.T., M.H., R.G.M.), Department of Biochemistry and Molecular Biology (R.G.M.), and Department of Medicine (R.G.M.), Indiana University School of Medicine, Indianapolis, Indiana 46202.
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Templin
authors.forename: Bernhard
authors.initials: B
authors.lastname: Maier
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Tersey
authors.forename: Masayuki
authors.initials: M
authors.lastname: Hatanaka
authors.forename: Raghavendra G
authors.initials: RG
authors.lastname: Mirmira
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D045744:Cell Line, Tumor; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics*; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D007515:Islets of Langerhans / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D011132:Polyribosomes / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D056811:Unfolded Protein Response / Q000235:genetics*
chemical_list: D020121:5' Untranslated Regions; D018398:Homeodomain Proteins; D053583:Interleukin-1beta; D012333:RNA, Messenger; D015534:Trans-Activators; D014409:Tumor Necrosis Factor-alpha; C084010:pancreatic and duodenal homeobox 1 protein; D007371:Interferon-gamma