Expression and regulation of mouse selenoprotein P transcript variants differing in non-coding RNA.
Selenoprotein P (Sepp1), a glycoprotein rich in selenium, is thought to function in selenium transport throughout the body. The sepp1 gene locus potentially produces three alternative transcripts that differ only in their 5' untranslated regions (5'UTRs) and not in their protein coding regions, as indicated by transcript information in genomic databases. Here we investigated the distribution, relative expression, and biological significance of these transcript variants. We confirmed the expression of Sepp1 transcript variants using PCR and sequencing. Using 5'-RACE, we identified multiple 5'-termini upstream from three different splice donor sites, and a single splice acceptor site for exon 2. We found regional and temporal changes in variant expression in select adult and neonate murine tissue and brain regions. Distribution of variants in heart and kidney varied with stage of development. Notably, the Sepp1b variant was localized specifically to the hippocampus in brain. Targeted silencing of individual variants using RNAi demonstrated the biological importance for all transcript variants in cell viability. Additionally, we determined that the Sepp1b variant is a specific target for the miR-7 microRNA by means of its unique 5'UTR structure. Our results emphasize the importance of non-coding transcript variations as a regulatory means for Sepp1 expression in different tissues and stages of development. The presence of a variant localized in the hippocampus and regulated by a microRNA may have implications for the known deficits in synaptic function caused by genetic deletion of Sepp1.
Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawai'I, Honolulu, HI, USA.
Andrea S T
AS
Dewing
Rachel H
RH
Rueli
Michael J
MJ
Robles
Elizabeth D
ED
Nguyen-Wu
Thomas
T
Zeyda
Marla J
MJ
Berry
Frederick P
FP
Bellinger
D017398:Alternative Splicing / Q000235:genetics*; D000818:Animals; D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D015870:Gene Expression; D017136:Ion Transport; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism; D020033:Protein Isoforms; D034622:RNA Interference; D022821:RNA Splice Sites; D034741:RNA, Small Interfering; D022661:RNA, Untranslated / Q000235:genetics*; D012643:Selenium / Q000378:metabolism; D051149:Selenoprotein P / Q000235:genetics* / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D020506:Untranslated Regions / Q000235:genetics
C549243:MIRN7 microRNA, mouse; D035683:MicroRNAs; D020033:Protein Isoforms; D022821:RNA Splice Sites; D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D051149:Selenoprotein P; D020506:Untranslated Regions; D012643:Selenium