The nuclear export signal-dependent localization of oligonucleopeptides enhances the inhibition of the protein expression from a gene transcribed in cytosol.
Upon endocytosis, most oligodeoxynucleotides (ODNs) accumulate in vesicular compartments; a tiny number of them cross the vesicle membrane, reach the cytosol and by passive diffusion enter the nucleus where they are entrapped. So far, the compartment in which an antisense ODN interacts with its mRNA target has not been precisely characterized. In an attempt to answer this question, ODN-peptides were designed with the aim of maintaining them in the cytosol. This has been achieved by a short peptide sequence called the nuclear export signal (NES). Upon microinjection, ODN-NES peptide conjugates were efficiently and rapidly exported from the nucleus to the cytosol whereas ODN-peptides containing an inactive NES were found to be located in the nucleus. The inhibitory activity of antisense ODN was tested in a system allowing the specific transcription of a luciferase reporter gene in the cytosol. Antisense propynylated ODN-NES peptide conjugates, directed against the luciferase gene, efficiently inhibited (75%) the cytosolic expression of luciferase whereas at the same concentration the peptide-free propynylated ODN or the propynylated ODN-peptides containing an inactive NES were nearly inactive.
Meunier
L
L
Glycobiologie, Centre de Biophysique Moléculaire, CNRS, Rue Charles Sadron, 45071 Orléans cedex 02, France.
Mayer
R
R
Monsigny
M
M
Roche
A C
AC
D000818:Animals; D002451:Cell Compartmentation; D002522:Chlorocebus aethiops; D003600:Cytosol; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014709:Vero Cells
D016376:Oligonucleotides, Antisense; D011506:Proteins; D012333:RNA, Messenger