Novel internucleotide 3'-NH-P(CH3)(O)-O-5' linkage. Oligo(deoxyribonucleoside methanephosphonamidates); synthesis, structure and hybridization properties.

Abstract:

:Diastereomeric dithymidine methanephosphonamidates (TnpmT) were synthesized by reaction of 3'-amino-3'-deoxythymidine with 3'- O -acetylthymidin-5-yl-methanephosphonochloridate. Separated dinucleotide TnpmT(fast) and TnpmT(slow) diastereomers were used as building blocks to prepare chimeric dodecathy-midylates, possessing one to four modified linkages, by means of phosphoramidite automated solid phase synthesis. As expected, the methanephosphonamidate internucleotide linkage is resistant to nuclease P1, snake venom PDE and 3'-exonuclease from human plasma. Degradation of dodecathymidylates possessing modified internucleotide linkages in alternate positions proved the 'hopping' properties of 3'-exonuclease. Oligo(deoxyribonucleotide methanephosphonamidates) were tested for their binding affinity to complementary oligomers in thermal denaturation experiments. All the oligomers showed lower binding affinity to DNA and RNA targets, however, oligomers originating from the TnpmT(fast) dimeric unit exhibited better hybridization properties than their diastereomeric TnpmT(slow) counterparts. A lowering of T m of approximately 2.4 degrees C (1.0-1.8 degrees C) was observed for each introduced TnpmT(fast) modification and 6.0 degrees C (4.2-5.0 degrees C) for each TnpmT(slow) modification in duplexes of modified dodecathymidylates with dA12(A12) oligomers. The oligo(deoxyribonucleoside methanephosphonamidate) designated F4, possessing four modified methanephosphonate linkages originating from the TnpmT(fast) diastereomeric unit, exhibits a tendency for triplex formation, as was demonstrated in thermal denaturation experiments with the d(A21C4T21) hairpin oligomer.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Nawrot B,Boczkowska M,Wójcik M,Sochacki M,Kazmierski S,Stec WJ

doi

10.1093/nar/26.11.2650

subject

Has Abstract

pub_date

1998-06-01 00:00:00

pages

2650-8

issue

11

eissn

0305-1048

issn

1362-4962

pii

gkb459

journal_volume

26

pub_type

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