PT  - JOURNAL ARTICLE
AU  - Pichon, Chantal
AU  - Arar, Khalil
AU  - Stewart, Alistair J.
AU  - Dodon, Madeleine Duc
AU  - Gazzolo, Louis
AU  - Courtoy, Pierre J.
AU  - Mayer, Roger
AU  - Monsigny, Michel
AU  - Roche, Annie-Claude
TI  - Intracellular Routing and Inhibitory Activity of Oligonucleopeptides Containing a KDEL Motif
DP  - 1997 Mar 01
TA  - Molecular Pharmacology
PG  - 431--438
VI  - 51
IP  - 3
4099  - http://molpharm.aspetjournals.org/content/51/3/431.short
4100  - http://molpharm.aspetjournals.org/content/51/3/431.full
SO  - Mol Pharmacol1997 Mar 01; 51
AB  - On internalization, oligonucleotides (ODN) remain mostly sequestered in endocytic compartments. To increase their delivery into the cytosol and/or nucleus, which contain their targets, we attempted to guide them into compartments containing the KDEL receptor. Antisense ODN, phosphodiester protected at both ends, that are complementary to the AUG initiation site of gagHIV-1mRNA (odn) were linked to a peptide ending with the Lys-Asp-Glu-Leu (KDEL) motif in a carboxyl-terminal position (odn-p-KDEL) or with the Lys-Asp-Glu-Ala (odn-p-KDEA) as a control. The effect of odn substitution with a peptide was examined with regard to its accumulation, subcellular location, and activity in HepG2 cells. Although odn-p-KDEL was internalized 4-fold less than the corresponding peptide-free odn, it was 5-fold more efficient in inhibitinggagHIV-1 gene expression in HepG2 cells. The internalization of odn-p-KDEA was as low as that of odn-p-KDEL, but its biological activity was lower, close to that of the peptide-free odn. On endocytosis at 37°, both conjugates as well as the peptide-free odn were found in a neutral environment. However, the substitution of an odn with a KDEL motif altered its intracellular trafficking; most of the odn-p-KDEL was found in the endoplasmic reticulum and in the intermediate compartment as identified by colabeling with either anti-ERGIC-53 or anti-KDEL receptor antibodies. Conversely, odn-p-KDEA and peptide-free odn were localized in vesicular compartments not labeled with these antibodies. In addition, pulse-chase experiments showed that odn-p-KDEL and odn-p-KDEA had a lower efflux than peptide-free odn. Therefore, the large increase in efficiency was due to the KDEL motif. The American Society for Pharmacology and Experimental Therapeutics