Synthesis and RNA polymerase incorporation of the degenerate ribonucleotide analogue rPTP.
The synthesis and enzymatic incorporation into RNA of the hydrogen bond degenerate nucleoside analogue 6-(beta-d-ribofuranosyl)-3, 4-dihydro-8H-pyrimido[4,5-c]-[1,2]oxazin-7-one (P) is described. The 5'-triphosphate of this analogue is readily incorporated by T3, T7 and SP6 RNA polymerases into RNA transcripts, being best incorporated in place of UTP, but also in place of CTP. When all the uridine residues in an HIV-1 TAR RNA transcript are replaced by P the transcript has similar characteristics to the wild-type TAR RNA, as demonstrated by similar melting temperatures and CD spectra. The P-substituted TAR transcript binds to the Tat peptide ADP-1 with only 4-fold lowered efficiency compared with wild-type TAR.
Moriyama
K
K
Gene Research Centre, Okayama University, Tsushima, Okayama 700, Japan, Nycomed Amersham plc, Amersham Laboratories, White Lion Road, Amersham, Buckinghamshire HP7 9LL, UK.
Negishi
K
K
Briggs
M S
MS
Smith
C L
CL
Hill
F
F
Churcher
M J
MJ
Brown
D M
DM
Loakes
D
D
D001483:Base Sequence; D002942:Circular Dichroism; D003570:Cytidine Triphosphate / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000737:chemistry; D012367:RNA, Viral / Q000096:biosynthesis* / Q000737:chemistry; D014544:Uridine Triphosphate / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism*; D014764:Viral Proteins
D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D003570:Cytidine Triphosphate; C070852:RNA polymerase SP6; C081591:bacteriophage T3 RNA polymerase; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D014544:Uridine Triphosphate