2'(3')-O-glycyl oligoribonucleotides with sequences of the 3'-terminus of glycyl-tRNA: chemical synthesis and properties in partial reactions of protein biosynthesis.
Specific syntheses of 2'(3')-O-aminoacyl oligoribonucleotides C-C-A-Gly (12), C-C-A(AcGly) (7), U-C-C-A-Gly (17), and C-U-C-C-A-Gly (19), which are the 3'-terminal sequences of Escherichia coli Gly-tRNA (or AcGly-tRNA, respectively) are described. Compounds 12, 17, and 19 were synthesized by employing the benzotriazolyl phosphotriester approach with the following protection groups on the components: benzoyl for the heterocyclic amino groups, 2-chlorophenyl group for internucleotide phosphate protection, dimethoxytrityl and levulinoyl groups for blocking of the 5'-hydroxyl, methoxytetrahydropyranyl group for protection of the 2'-hydroxyl functions, and N-(benzyloxycarbonyl)orthoglycinate as the masked aminoacyl group simultaneously protecting the 2',3'-cis diol group of the 3'-terminal adenosine moiety. The fully protected tri-, tetra-, and pentanucleotides were obtained via 5'-extension of di- and trinucleotide blocks after prior selective removal of the 5'-O-levulinoyl group with hydrazine. The blocked derivatives 11, 16, and 18 were totally deprotected by reactions with NH4OH, H+, and H2/Pd to yield the target compounds 12, 17, and 19 in good yields. C-C-A(AcGly) (7) was synthesized according to a stepwise procedure via activation of preformed diesters with (mesitylenesulfonyl)tetrazole. C-C-A-Gly (12), U-C-C-A-Gly (17), and C-U-C-C-A-Gly (19) were all acceptor substrates in the peptidyltransferase reaction with the Ac-Phe-tRNA-70S ribosome-poly(U) system. All three models also promoted EF-Tu-70S ribosome GTP hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS)
Scalfi-Happ
C
C
Department of Chemistry, Michigan Cancer Foundation, Detroit 48201.
Happ
E
E
Ghag
S
S
Chládek
S
S
D001483:Base Sequence; D006160:Guanosine Triphosphate / Q000378:metabolism; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D009843:Oligoribonucleotides / Q000138:chemical synthesis*; D010444:Peptide Elongation Factor Tu / Q000378:metabolism; D014176:Protein Biosynthesis; D012346:RNA, Transfer, Amino Acyl / Q000138:chemical synthesis* / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D007202:Indicators and Reagents; D009843:Oligoribonucleotides; D012346:RNA, Transfer, Amino Acyl; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu