Multiple initiation sites of DNA replication flanking the origin region of λdv genome

Toshiki Tsurimoto, K. Matsubara

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11 Citations (Scopus)

Abstract

Early replicative intermediates of λdv plasmid were prepared by an in vitro replication system in the presence of 2',3'-dideoxycytidine 5'-triphosphate, an inhibitor of DNA chain elongation. Short-chain DNAs produced from regions near the replication origin were purified from the intermediates. A fraction of the DNAs was covalently linked to primer RNA. The transition sites from primer RNA to DNA synthesis were mapped along the nucleotide sequence of the genome, by eliminating the RNA by alkaline hydrolysis and labeling the freshly exposed 5' ends of DNA with 32P. The transition sites were found to be located on both sides of the ori region, which includes four 19-base-pair repeats where one of the λ specific initiator proteins, O, binds. No transition arose within the ori region. The transition sites are multiple on both sides of the ori region and are clustered in one of the two strands in such a way that DNA syntheses from the two sides converge. The frequency of the 'leftward' DNA synthesis is several times higher than that of 'rightward' synthesis, reflecting the asymmetric bidirectional replication of λdv DNA.

Original languageEnglish
Pages (from-to)7402-7406
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number23 I
DOIs
Publication statusPublished - Jan 1 1984

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Replication Origin
DNA Replication
Genome
DNA
Base Pairing
Hydrolysis
Plasmids
RNA

All Science Journal Classification (ASJC) codes

  • General

Cite this

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abstract = "Early replicative intermediates of λdv plasmid were prepared by an in vitro replication system in the presence of 2',3'-dideoxycytidine 5'-triphosphate, an inhibitor of DNA chain elongation. Short-chain DNAs produced from regions near the replication origin were purified from the intermediates. A fraction of the DNAs was covalently linked to primer RNA. The transition sites from primer RNA to DNA synthesis were mapped along the nucleotide sequence of the genome, by eliminating the RNA by alkaline hydrolysis and labeling the freshly exposed 5' ends of DNA with 32P. The transition sites were found to be located on both sides of the ori region, which includes four 19-base-pair repeats where one of the λ specific initiator proteins, O, binds. No transition arose within the ori region. The transition sites are multiple on both sides of the ori region and are clustered in one of the two strands in such a way that DNA syntheses from the two sides converge. The frequency of the 'leftward' DNA synthesis is several times higher than that of 'rightward' synthesis, reflecting the asymmetric bidirectional replication of λdv DNA.",
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AU - Matsubara, K.

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N2 - Early replicative intermediates of λdv plasmid were prepared by an in vitro replication system in the presence of 2',3'-dideoxycytidine 5'-triphosphate, an inhibitor of DNA chain elongation. Short-chain DNAs produced from regions near the replication origin were purified from the intermediates. A fraction of the DNAs was covalently linked to primer RNA. The transition sites from primer RNA to DNA synthesis were mapped along the nucleotide sequence of the genome, by eliminating the RNA by alkaline hydrolysis and labeling the freshly exposed 5' ends of DNA with 32P. The transition sites were found to be located on both sides of the ori region, which includes four 19-base-pair repeats where one of the λ specific initiator proteins, O, binds. No transition arose within the ori region. The transition sites are multiple on both sides of the ori region and are clustered in one of the two strands in such a way that DNA syntheses from the two sides converge. The frequency of the 'leftward' DNA synthesis is several times higher than that of 'rightward' synthesis, reflecting the asymmetric bidirectional replication of λdv DNA.

AB - Early replicative intermediates of λdv plasmid were prepared by an in vitro replication system in the presence of 2',3'-dideoxycytidine 5'-triphosphate, an inhibitor of DNA chain elongation. Short-chain DNAs produced from regions near the replication origin were purified from the intermediates. A fraction of the DNAs was covalently linked to primer RNA. The transition sites from primer RNA to DNA synthesis were mapped along the nucleotide sequence of the genome, by eliminating the RNA by alkaline hydrolysis and labeling the freshly exposed 5' ends of DNA with 32P. The transition sites were found to be located on both sides of the ori region, which includes four 19-base-pair repeats where one of the λ specific initiator proteins, O, binds. No transition arose within the ori region. The transition sites are multiple on both sides of the ori region and are clustered in one of the two strands in such a way that DNA syntheses from the two sides converge. The frequency of the 'leftward' DNA synthesis is several times higher than that of 'rightward' synthesis, reflecting the asymmetric bidirectional replication of λdv DNA.

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