We have used site-specifically spin-labeled polyribonucleotides to extract motional information for single-and double-stranded polyribonucleotides. The spin-label is attached to either position 4 or 5 of an uridine analogue, and in the latter position the label is linked through various tethers to the base.
The synthesis of one C(4)-spin-laheled uridine-S'-diphosphate (C(4)-UDP and two nitroxide-containing 2'-deoxyuridine-S'-triphosphate (dUTP) analogs are reported. The C(4)-UDP derivative was incorporated into copolymers by polynucleotide phosphoxylase (PNPase); one of' the two dUTP analogs, substituted at position C(4), was a good substrate for TdT, whereas the other one, substituted at position C(5), served as substrate for E. coii DNA polymerase (Pol 1).Introduction. ~ The application of the spin-labeled nucleic acids to investigate problems of biological interest has been well established in this as well as other laboratories [ 1-41. Of great interest is the synthesis of spin-labeled ribo-or deoxyribonucleotides which can serve as substrates for enzymes such as PNPase'), TdT, and Pol I in order to prepare enzymatically spin-labeled nucleic acids. The former two enzymes have no template requirement and were used in this laboratory to prepare a variety of RNA and DNA homopolymers containing different amounts of enzymatically incorporated spinlabeled ribo-or deoxyribonucleotides [5] [6]. Pol I, on the other hand, has a template requirement and is well suited to make template specific spin-labeled polynucleotides.We wish to report here the synthesis of the three novel spin-labeled nucleotides pplss4U (2), pppls4dU (6), and pppDUAT (12). pplss4U and pppls4dU served as substrates for PNPase and TdT, respectively, and pppDUAT was readily incorporated by Pol I. The biophysical properties of (Iss4U,U),, (ls4dU,dT), and (DUAT,dT), were described in [7].
Die Synthese von Uridin‐diphosphat‐Analogen, die am C‐5 (V) bzw. am (C4) (VII) eine Spin‐Marker‐Gruppierung tragen, sowie die enzymatische Inkorporierung in Ribonucleinsäuren (VIII) [→(IX) bzw. (X) (Copolymere mit Uridin bzw. Cytidin)] werden beschrieben.
The synthesis of uridine‐5′‐diphosphate analogs, spin‐labeled either at C(4) or C(5) is reported as well as their enzymatic incorporation into ribonucleic acids, some of which had previously been shown to be potent interferon inducers upon annealing with poly (inosinic acid). Also, the synthesis of spin‐labeled poly (cytidylic acid) obtained by chemical acylation is presented.
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