Extension of RNA molecules with DNA. An approach to the study of termination sequences recognised by RNA polymerase.

Sequences in DNA, which act as signals for the termination of transcription, may lie wholly or partially beyond the distal end of the DNA region actually transcribed into RNA. In a hybrid formed between an intact RNA molecule and its DNA complement, any such untranscribed signal should lie in the region of single-stranded DNA close to the 3′-hydroxyl terminus of the RNA. We show here that hybrids of this type, formed on circular bacteriophage DNA, act as efficient primers for Escherichia coli DNA polymerase I and T4-induced DNA polymerase, under the normal conditions for DNA synthesis in vitro. The DNA product is covalently linked to the 3′-end of the RNA, and co-purifies with it in CsCl equilibrium centrifugation. Priming by RNA is readily detectable even when DNA primers are also present. These and other features of the reaction suggest that it might form the basis of a general method for the study of DNA sequences beyond the termini of RNA chains.

[1]  W. Keller.  RNA-primed DNA synthesis in vitro. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[2]  P. T. Englund,et al.  The 3'-terminal nucleotide sequences of T7 DNA. , 1972, Journal of molecular biology.

[3]  R. Schekman,et al.  RNA synthesis initiates in vitro conversion of M13 DNA to its replicative form. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[4]  R. Matson,et al.  Membrane stability (thermal) and nature of fatty acids in yeast cells. , 1972, Biochemical and Biophysical Research Communications - BBRC.

[5]  R. Wells,et al.  Comparison of some reactions catalyzed by deoxyribonucleic acid polymerase from avian myeloblastosis virus, Escherichia coli, and Micrococcus luteus. , 1972, Biochemistry.

[6]  J. Hurwitz,et al.  RNA-Dependent DNA Polymerase Activity of RNA Tumor Viruses I. Directing Influence of DNA in the Reaction , 1972, Journal of virology.

[7]  R. Schekman,et al.  A possible role for RNA polymerase in the initiation of M13 DNA synthesis. , 1971, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. Darnell,et al.  Polyadenylic Acid Sequences: Role in Conversion of Nuclear RNA into Messenger RNA , 1971, Science.

[9]  D. Baltimore,et al.  Covalently linked RNA-DNA molecule as initial product of RNA tumour virus DNA polymerase. , 1971, Nature: New biology.

[10]  P. T. Englund,et al.  The initial step of in vitro synthesis of deoxyribonucleic acid by the T4 deoxyribonucleic acid polymerase. , 1971, The Journal of biological chemistry.

[11]  N. Pace,et al.  Primary structural relationship of p16 to m16 ribosomal RNA. , 1971, Nature: New biology.

[12]  C. Mitchell,et al.  Persistence of Morphine in Central Nervous System of Rats after a Single Injection and its Bearing on Tolerance , 1971, Nature.

[13]  R. Wu,et al.  Nucleotide sequence analysis of DNA. II. Complete nucleotide sequence of the cohesive ends of bacteriophage lambda DNA. , 1971, Journal of molecular biology.

[14]  W. S. Kelley,et al.  Purification of an Altered DNA Polymerase from an E. coli Strain with a pol Mutation , 1971, Nature.

[15]  U. Maitra.  The identification of 3'-hydroxyl nucleoside terminus of RNA chains released in the DNA-dependent RNA polymerase reaction. , 1970, Biochemical and Biophysical Research Communications - BBRC.

[16]  L. Cavalieri,et al.  RNA as a template with E. coli DNA polymerase. , 1970, Biochemical and biophysical research communications.

[17]  R. Wells,et al.  Micrococcus luteus deoxyribonucleic acid polymerase. Studies of the enzymic reaction and properties of the deoxyribonucleic acid product. , 1970, The Journal of biological chemistry.

[18]  J. Richardson.  Rates of bacteriophage T4 RNA chain growth in vitro. , 1970, Journal of molecular biology.

[19]  B. Alberts,et al.  Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. , 1970, Virology.

[20]  M. Chamberlin.  Transcription 1970: A Summary , 1970 .

[21]  Jeffrey W. Roberts.  Termination Factor for RNA Synthesis , 1969, Nature.

[22]  R. Burgess.  A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase. , 1969, The Journal of biological chemistry.

[23]  I. Barrai,et al.  Dependence of the buoyant density of single-stranded DNA on base composition. , 1969, Journal of molecular biology.

[24]  R. Symons.  Preparation of [alpha-32P]nucleoside and deoxynucleoside 5'-triphosphates from 32Pi and protected and unprotected nucleosides. , 1969, Biochimica et Biophysica Acta.

[25]  R. Schekman,et al.  Replication of bacteriophage M13. I. Sedimentation analysis of crude lysates of M13-infected bacteria. , 1969, Biochimica et biophysica acta.

[26]  A. Kornberg.  Active center of DNA polymerase. , 1969, Science.

[27]  J. Devinny,et al.  The use of an ethidium analogue in the dye-buoyant density procedure for the isolation of closed circular DNA: the variation of the superhelix density of mitochondrial DNA. , 1969, Proceedings of the National Academy of Sciences of the United States of America.

[28]  J. Dahlberg.  Terminal Sequences of Bacteriophage RNAs , 1968, Nature.

[29]  I. Lehman,et al.  An in vitro transversion by a mutationally altered T4-induced DNA polymerase. , 1968, Journal of molecular biology.

[30]  H. Hoffmann-Berling,et al.  Endonuclease-I-deficient and ribonuclease I-deficient Escherichia coli mutants. , 1968, Journal of molecular biology.

[31]  A. Kornberg,et al.  Enzymatic synthesis of deoxyribonucleic acid. XXV. Purification and properties of deoxyribonucleic acid polymerase induced by infection with phage T4. , 1968, The Journal of biological chemistry.

[32]  M. Riley,et al.  Physical and chemical characterization of two- and three-stranded adenine-thymine and adenine-uracil homopolymer complexes. , 1966, Journal of molecular biology.

[33]  N. Zinder,et al.  HOST-DEPENDENT MUTANTS OF THE BACTERIOPHAGE F2. I. ISOLATION AND PRELIMINARY CLASSIFICATION. , 1964, Virology.

[34]  I. Lehman,et al.  THE DEOXYRIBONUCLEASES OF ESCHERICHIA COLI. IV. AN EXONUCLEASE ACTIVITY PRESENT IN PURIFIED PREPARATIONS OF DEOXYRIBONUCLEIC ACID POLYMERASE. , 1964, The Journal of biological chemistry.

[35]  A. Kornberg,et al.  ENZYMATIC SYNTHESIS OF DEOXYRIBONUCLEIC ACID. XV. PURIFICATION AND PROPERTIES OF A POLYMERASE FROM BACILLUS SUBTILIS. , 1964, The Journal of biological chemistry.

[36]  S. Lee-Huang,et al.  POLYRIBONUCLEOTIDES AS TEMPLATES FOR POLYDEOXYRIBONUCLEOTIDES. , 1963, Proceedings of the National Academy of Sciences of the United States of America.

[37]  M. Chamberlin,et al.  DEOXYRIBONUCLEIC ACID-DIRECTED SYNTHESIS OF RIBONUCLEIC ACID BY AN ENZYME FROM ESCHERICHIA COLI , 1962 .