The use of 5'-phospho-2 deoxyribocytidylylriboadenosine as a facile route to chemical aminoacylation of tRNA.
Methodology is described for the synthesis and chemical aminoacylation of the hybrid dinucleotide 5'-phospho-2'-deoxyribocytidylylriboadenosine (pdCpA). Ligation of aminoacylated pdCpA to a truncated amber suppressor tRNACUA (-CA) using T4 RNA ligase generates an aminoacylated suppressor tRNA which can be used for site-specific incorporation of unnatural amino acids into proteins. Both the ligation and in vitro suppression efficiencies are the same when either pCpA or pdCpA is used. The use of deoxycytidine simplifies the chemistry involved in the synthesis of the dinucleotide pCpA. In addition, these results demonstrate that ribocytidine is not required for recognition of the aminoacylated tRNA during protein synthesis.
Robertson
S A
SA
Department of Chemistry, University of California, Berkeley 94720.
Noren
C J
CJ
Anthony-Cahill
S J
SJ
Griffith
M C
MC
Schultz
P G
PG
D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D015226:Dinucleoside Phosphates; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D015394:Molecular Structure; D012343:RNA, Transfer; D012346:RNA, Transfer, Amino Acyl / Q000138:chemical synthesis*; D013053:Spectrophotometry; D013489:Suppression, Genetic
D015226:Dinucleoside Phosphates; D007202:Indicators and Reagents; D012346:RNA, Transfer, Amino Acyl; C062195:5'-phospho-2'-deoxyribocytidylylriboadenosine; D012343:RNA, Transfer