IRESite record type: plasmid_with_promoter_and_putative_IRES_translationally_characterized
The shape of the nucleic acid molecule translated: linear
The quality of the mRNA/+RNA sequence: 3UTR_possibly_incomplete
The mRNA/+RNA description:
Putative in vivo CMV promoter-derived transcript produced from monocistronic plasmid
pHIAP2(1073-1222)(CUA1147)/CAT which contains the part of human c-IAP1 5' UTR (nt from -150 to nt -1 of the
original sequence) mRNA, where the authentic initiation CUG (-76 to -74) codon of c-IAP1 upstream ORF (uORF)
was substituted for CUA, ahead of chloramphenicol acetyltransferase reporter gene.
The sequence ends at its 3'-end right after the poly(A) signal from BGH mRNA and thus the 3'-UTR might be
slightly wrong.
The mRNA/+RNA sequence represented in the +DNA notation:
Credibility of mRNA sequence: reverse_engineered_sequence_and_should_match_experiment_except_3UTR
The name of the promoter used to express this mRNA: CMV
Aliases of the plasmid name:
Alias: HIAP2(1073-1222)(CUA1147)CAT
Description of the plasmid (facultative for promoter-less plasmid records): Plasmid containing part of human c-IAP1 5' UTR (nt from -150 to nt -1 of the original sequence) ahead of
chloramphenicol acetyl transferase reporter gene. The authentic initiation CTG (-76 to -74) codon of c-IAP1
upstream ORF (uORF) was substituted for CTA.
The in vivo produced transcripts are heterogeneous (due to any of promoter?/splicing?/cleavage?/breakage?): not tested
The in vivo produced heterogeneous transcripts occur due to alternative splicing: not tested
A promoter reported in cDNA corresponding to IRES sequence: not tested
The abbreviated name of the donor gene or virus from which this IRES was excised and inserted into the plasmid: c-IAP1
The IRES absolute position (the range includes START and STOP codons or their equivalents): 148-297
How IRES boundaries were determined: experimentally_determined
The sequence of IRES region aligned to its secondary structure (if available):
Remarks:
c-IAP1_-150/-1_CUA1147 IRES represents part of the human c-IAP1 5' UTR (nt from -150 to nt -1) where the
authentic initiation CTG (-76 to -74) codon of c-IAP1 upstream ORF (uORF) was substituted for CTA.
The relative translation efficiency in % of this IRES: 100.000
Name of the plasmid used as the negative control. pHIAP2(1073-1222)/CAT
IRESite Id of the plasmid used as negative control. 536
The relative translation efficiency in % of the negative control: 4.000
The size (length) of intercistronic region in the negative control: 0
The effect of 5'-cap analogs on translation: not tested
Rapamycin affects translation: not tested
Type of RNA subject to translation: endogenous_nuclear_RNA_Pol_II_transcript
Remarks:
In the monocistronic CAT reporter plasmids where the 5' UTR of c-IAP1 (with or without the mutation in the
uORF-initiating CUG) is fused with the CAT gene is the translation of CAT from the 5' UTR containing
construct, but not in the absence of the 5' UTR, enhanced in drug-treated (with thapsigargin or tunicamycin)
cells. This observation paralleled the behavior of the endogenous c-IAP1 in drug-treated cells, suggesting
that the induction of c-IAP1 after endoplasmic reticulum (ER) stress is due to the translation up-regulation
that is mediated by the 5' UTR. Surprisingly, the CUG to CUA mutation that prevents the translation of uORF
had no effect on the induction of CAT after ER stress. Therefore, although the 5' UTR mediates induction of
c-IAP1 in response to ER stress, this induction is independent of the regulatory uORF (Warnakulasuriyarachchi
et al., 2004).
Translational data are derived from Fig. 4B.