Stress-sensitive regulation of IFRD1 mRNA decay is mediated by an upstream open reading frame.
In this report, we demonstrate that cellular stress regulates expression of IFRD1 by a post-transcriptional control mechanism. IFRD1 mRNA and protein are elevated in tunicamycin-treated human kidney epithelial cells via stabilization of the mRNA. IFRD1 mRNA instability in resting cells requires translation of an upstream open reading frame (ORF) that represses translation of the major ORF. During stress response, the mRNA is stabilized via inhibition of translational initiation mediated by phosphorylated eIF2alpha. Translation of the major ORF of IFRD1 involves both leaky scanning at the upstream AUG codon and re-initiation at the major AUG codon and is not altered during stress. Finally, the instability mechanism depends upon UPF1, suggesting that it is related to nonsense-mediated decay. Importantly, the sequence and length of the upstream ORF are critical but do not need to code for a specific peptide. Moreover the sequence environment of the upstream ORF termination site is not an essential feature of instability. These features of decay collectively define a distinct upstream ORF-mediated instability mechanism whereby cellular stress can modulate specific gene expression through alteration of mRNA half-life.
Zhao
Chenyang
C
Department of Immunology, Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
Datta
Shyamasree
S
Mandal
Palash
P
Xu
Shuqing
S
Hamilton
Thomas
T
D001665:Binding Sites; D003062:Codon; D004847:Epithelial Cells / Q000166:cytology; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism / Q000502:physiology*; D007668:Kidney / Q000166:cytology; D008954:Models, Biological; D016366:Open Reading Frames; D010766:Phosphorylation; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*; D013314:Stress, Mechanical
D003062:Codon; D015852:Eukaryotic Initiation Factor-2; C490045:IFRD1 protein, human; D017874:Immediate-Early Proteins; D012333:RNA, Messenger