The use of oligonucleotide probes containing 2'-deoxy-2'-fluoronucleosides for regiospecific cleavage of RNA by RNase H from Escherichia coli.
Protected 2'-deoxy-2'-fluorouridine and 2'-deoxy-2'-fluorocytidine suitable for incorporation into oligonucleotides via the phosphoramidite approach have been prepared. Five modified and two unmodified oligonucleotides have been synthesized to investigate the regiospecific cleavage of a 5S RNA from Escherichia coli by RNase H. In order to show whether the modified oligonucleotides are able to hybridize with the RNA the physico-chemical properties (melting curves, CD spectra) of analogous DNA/oligodeoxyribonucleotide duplexes have been examined. The modified oligonucleotides are shown to form stable duplexes with a DNA-matrix which exist in an A-like form. Two of the modified probes containing four 2'-deoxy-2'-fluorocytidines or two 2'-deoxy-2'-fluorouridines direct the splitting by RNase H of only one phosphodiester bond of the RNA.
Schmidt
S
S
Humboldt University, Institute of Bioorganic Chemistry, Berlin, Germany.
Niemann
A
A
Krynetskaya
N F
NF
Oretskaya
T S
TS
Metelev
V G
VG
Suchomlinov
V V
VV
Shabarova
Z A
ZA
Cech
D
D
D001483:Base Sequence; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D005467:Floxuridine / Q000378:metabolism*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes / Q000378:metabolism*; D012329:RNA, Bacterial / Q000378:metabolism*; D012341:RNA, Ribosomal, 5S / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism*
D015345:Oligonucleotide Probes; D012329:RNA, Bacterial; D012341:RNA, Ribosomal, 5S; D012360:RNA, Transfer, Phe; D005467:Floxuridine; D003841:Deoxycytidine; C037289:2'fluoro-2'-deoxyuridine; D016914:Ribonuclease H; C007251:2'-fluoro-2'-deoxycytidine