Effects of 5' untranslated region diversity on the posttranscriptional regulation of the human reduced folate carrier.
The human RFC (hRFC) gene is regulated by five major 5' non-coding exons, characterized by alternate transcription start sites and splice forms. The result is up to 14 hRFC transcripts for which different 5' untranslated regions (UTRs) are fused to a common coding sequence. By in vitro translation assays with hRFC constructs corresponding to the major transcript forms, most of the forms were translated poorly. Upon expression of the 5'UTR-hRFC constructs in hRFC-null HeLa cells, a range of steady state hRFC proteins and transcripts were detected that reflected relative transcript stabilities and, to a lesser extent, translation efficiencies. Transcripts including 5' UTRs derived from non-coding exon A encoded a modified hRFC protein translated from an upstream initiation site. When this modified hRFC protein was expressed in hRFC-null K562 cells, there were only minor differences in surface targeting, stability, or transport function from wild type hRFC. Our results demonstrate an important role for posttranscriptional determinants of cellular hRFC levels and activity.
Payton
Scott G
SG
Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Haska
Christina L
CL
Flatley
Robin M
RM
Ge
Yubin
Y
Matherly
Larry H
LH
D020121:5' Untranslated Regions / Q000737:chemistry*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002478:Cells, Cultured; D005091:Exons; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D007700:Kinetics; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D058978:Reduced Folate Carrier Protein; D014162:Transfection
D020121:5' Untranslated Regions; D026901:Membrane Transport Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D058978:Reduced Folate Carrier Protein; C548699:SLC19A1 protein, human