The nucleic acid data:
IRESite Id: 535 Version: 3
Originaly submitted by: Václav Vopálenský
Reviewed by: Martin Mokrejš Last change: 2008-10-27 23:46:47
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_incomplete
The mRNA/+RNA description: 
Putative in vivo CMV promoter-derived transcript produced from monocistronic plasmid pHIAP2(990-1222)/CAT
which contains the part of human c-IAP1 5' UTR (nt from -233 to nt -1 of the original sequence) mRNA 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_maybe_except_3UTR
The name of the plasmid:
pHIAP2(990-1222)/CAT
The name of the promoter used to express this mRNA:
  CMV
Aliases of the plasmid name:
Alias: HIAP2(990-1222)CAT
Description of the plasmid (facultative for promoter-less plasmid records):
Plasmid containing part of human c-IAP1 5' UTR (nt from -233 to nt -1 of the original sequence) ahead of chloramphenicol acetyltransferase reporter gene.
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(990-1222)/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 23:46:47
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):
  387-1046
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: 1 Last change: 2008-10-23 12:57:52
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The IRES name:
  c-IAP_-233/-1
The functional status of IRES:
  functional
The IRES absolute position (the range includes START and STOP codons or their equivalents):
  148-380
How IRES boundaries were determined:
experimentally_determined
The sequence of IRES region aligned to its secondary structure (if available):


Remarks:
c-IAP1_-233/-1 IRES represents part of the human c-IAP1 5' UTR (nt from -233 to nt -1).
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: 583
Version: 3 Last change: 2008-10-23 12:57:52
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:
  1.000
Name of IRES used as the positive control:
  XIAP
Name of the plasmid used as the positive control.
pXIAP(1-1076)/CAT
The relative translation efficiency in % of the positive control:
  100.000
The size (length) of intercistronic region in the positive 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:
The c-IAP1 5' UTR contains an upstream ORF (uORF) that prevents translation of the c-IAP1 ORF under normal
conditions (Warnakulasuriyarachchi et al., 2003) (this case of translation experiment).
Subsequent investigations revealed an IRES element at position -149 to position -1 in the c-IAP1 5' UTR that
mediates c-IAP1 translation following endoplasmic reticulum (ER) stress (treatment with etoposide or with
sodium arsenate (Van Eden et al., 2004) or treatment with thapsigargin (an inhibitor of ER calcium pump) or
tunicamycin (an inhibitor of protein glycosylation) (Warnakulasuriyarachchi et al., 2004)). Thus, expression
of c-IAP1 is regulated by a cap-independent translation initiation mechanism following cell stress. ER stress
causes cleavage of the eIF-4G family member p97/DAP5/NAT1 from its p97 form to a p86 form, and the p86 form
induces the activity of the c-IAP1 IRES resulting in an increase in c-IAP1 protein levels. However, direct
binding of the p86 form of p97/DAP5/NAT1 to c-IAP1 IRES RNA has not yet been demonstrated, and therefore p86
may exert its effect on c-IAP1 IRES activity either directly or via a downstream ITAF that interacts with the
c-IAP1 IRES.

Translational data are derived from Fig. 3A.
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
Last change to the database: 2015-04-16 16:45:23 GMT+1