Letter: Terminal deoxynucleotidyltransferase in murine myelomas.
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Biomedical subjects
Publications and source records attributed to F Chapeville.
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The coding properties of RNA transcribed in vitro by purified vaccinia cores have been investigated using Krebs ascites tumor cells, L cells, and reticulocyte lysates. Six to 10 proteins synthesized in vitro are separated on polyacrylamide gels by electrophoresis in the presence of sodium dodecyl sulfate. Their molecular weights vary from 10,000 to 44,000. The electrophoretic behavior of these proteins is similar to that of early proteins isolated from infected L cells. The tryptic peptide analysis of one of these proteins indicates similarity in amino acid sequences. These results show fidelity of both in vitro transcription and molecular weight above 44,000 are synthesized in vitro does not seem due to a competition between 12S mRNA synthesized in excess and RNA of a higher sedimentation coefficient present in a lower amount.
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A DNA polymerase purified from extracts of chick embryo catalyzes the synthesis of poly(dT) in the presence of poly(rA) or poly(dA) as template and poly(dT) or oligo(dT) as primer. Oligo- or polyribonucleotides are relatively ineffective initiators of polydeoxynucleotide polymerization. Using activated DNA as template, the enzyme catalyzes a repair-like reaction that resembles that catalyzed by RNA-directed DNA polymerase of RNA tumor viruses and Escherichia coli DNA polymerase II. With natural templates containing poly(rA) sequences, such as avian myeloblastosis virus RNA or rabbit globin mRNA, and oligo(dT) as initiator, principally poly(dT) is synthesized. Consequently, the enzyme differs from the RNA-directed DNA polymerase associated with RNA tumor viruses by its incapacity to transcribe heteropolymeric regions of RNA templates. The possible role of the enzyme in repair of DNA during transcription is discussed.
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Turnip yellow mosaic virus (TYMV) RNA treated with snake venom phosphodiesterase accepts cytidine 5'-monophosphate and adenosine 5'-monophosphate (AMP) when it is incubated in the presence of cytidine 5'-triphosphate (CTP), adenosine 5'-triphosphate, and Escherichia coli transfer RNA nucleotidyltransferase; untreated TYMV RNA accepts only AMP. When alpha (32)PCTP was used for terminal labeling, the nearest neighbor analyses and the anallyses after action of various nucleases showed that the sequence of five nucleotides at the 3' end of TYMV RNA is: pGpCpApCpC. A nuclease present in commerical preparations of snake venom phosphodiesterase leads to the fragmentation of TYMV RNA, the 3' end of which is found in a fragment having a sedimentation constant close to 5s.
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