Synthesis and binding properties of oligonucleotides carrying nuclear localization sequences.
The synthesis of oligonucleotides carrying nuclear localization peptide sequences is described using two strategies: first, oligonucleotides carrying a thiol group at the 5' end were reacted with maleimido peptides; second, peptide and oligonucleotide were prepared stepwise on the same support, yielding oligonucleotide-3'-peptide conjugates. This second approach was thoroughly studied. Using amino acids and small peptides as model compounds, some side reactions were analyzed, detected, and minimized. Oligonucleotides complementary to Ha-ras gene and carrying nuclear localization peptides at the 3' and 5' ends were prepared. Melting temperature studies showed that duplexes containing nuclear localization peptides were more stable than duplexes with unmodified oligonucleotides. Moreover, oligonucleotide-peptide conjugates maintain a good mismatch discrimination when they bind to their target RNA.
de La Torre
B G
BG
Centro de Investigación y Desarrollo, C.S.I.C. Jordi Girona 18-26, E-08034 Barcelona, Spain.
Albericio
F
F
Saison-Behmoaras
E
E
Bachi
A
A
Eritja
R
R
D000595:Amino Acid Sequence; D000641:Ammonia; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D004355:Drug Stability; D011905:Genes, ras; D007930:Leucine; D008301:Maleimides / Q000737:chemistry; D019913:Nuclear Localization Signals; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000138:chemical synthesis*; D009842:Oligopeptides / Q000737:chemistry*; D010649:Phenylalanine; D012313:RNA / Q000378:metabolism; D013053:Spectrophotometry; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014633:Valine
D008301:Maleimides; D019913:Nuclear Localization Signals; D009841:Oligonucleotides; D009842:Oligopeptides; C402304:SV40 nuclear localization peptide; D013438:Sulfhydryl Compounds; D010649:Phenylalanine; D012313:RNA; D000641:Ammonia; D007930:Leucine; D014633:Valine