The nucleic acid data:
IRESite Id: 539 Version: 4
Originaly submitted by: Václav Vopálenský
Reviewed by: Martin Mokrejš Last change: 2008-10-27 23:36:27
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 plasmid:
pHIAP2(1073-1222)(CUA1147)/CAT
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 origin of IRES in the plasmid:
  nuclear
The donor organism of the IRES segment:
Homo sapiens Jurkat
The DNA sequence of the plasmid in (+) orientation annotated by its secondary structure:


GenBank formatted file with annotated plasmid sequence hyperlinked from vector image map:
pHIAP2(1073-1222)(CUA1147)/CAT.jpg
The total number of notable open-reading frames (ORFs):
  1
Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:
ORF
ORF position:   1
Version: 1 Last change: 2008-10-27 23:36:27
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The abbreviated name of this ORF/gene:
CAT
The description of the protein encoded in this ORF:
chloramphenicol acetyltransferase
The translational frameshift (ribosome slippage) involved:
  0
The ribosome read-through involved:
  no
The alternative forms of this protein occur by the alternative initiation of translation:
  not tested
The ORF absolute position (the base range includes START and STOP codons or their equivalents):
  304-963
Citations:
Warnakulasuriyarachchi D., Ungureanu N. H., Holcik M. (2003) The translation of an antiapoptotic protein HIAP2 is regulated by an upstream open reading frame. Cell. Death. Differ. 10(8):899-904
IRESs:
IRES:
Version: 2 Last change: 2008-10-23 13:27:14
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The IRES name:
  c-IAP1_-150/-1_CUA1147
The functional status of IRES:
  functional
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.
Citations:
Warnakulasuriyarachchi D., Ungureanu N. H., Holcik M. (2003) The translation of an antiapoptotic protein HIAP2 is regulated by an upstream open reading frame. Cell. Death. Differ. 10(8):899-904
The translation experiments:
Translation results:
IRESite Translation Id: 587
Version: 7 Last change: 2008-10-23 13:27:14
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The translation method used to study IRES function:
in vivo
The organism used for translation:
Homo sapiens HEK 293T (ATCC 293tsA1609neo)
The temperature (in degrees of Celsia):
37
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.
Citations:
Warnakulasuriyarachchi D., Ungureanu N. H., Holcik M. (2003) The translation of an antiapoptotic protein HIAP2 is regulated by an upstream open reading frame. Cell. Death. Differ. 10(8):899-904
Warnakulasuriyarachchi D., Cerquozzi S., Cheung H. H., Holcik M. (2004) Translational induction of the inhibitor of apoptosis protein HIAP2 during endoplasmic reticulum stress attenuates cell death and is mediated via an inducible internal ribosome entry site element. J. Biol. Chem. 279(17):17148-17157
Last change to the database: 2015-04-16 16:45:23 GMT+1