Start sites of bidirectional DNA synthesis at the human lamin B2 origin |
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Authors: | Abdurashidova G Deganuto M Klima R Riva S Biamonti G Giacca M Falaschi A |
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Institution: | Molecular Biology and Molecular Medicine Units, International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34012 Trieste, Italy. |
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Abstract: | The initiation sites of bidirectional synthesis at the DNA replication origin located at the 3' end of the human lamin B2 gene were investigated. RNA-primed nascent DNA molecules were subjected to second-strand synthesis with appropriate primers, amplified by ligation-mediated polymerase chain reaction, and size fractionated. Evidence for precise start sites was obtained. Exploration of close to 1 kilobase, coupled to inhibition of Okazaki fragment synthesis, demonstrates that the leading strands initiate at precise nucleotides on either helix, overlapping by three base pairs, within the area bound to a protein complex possibly analogous to the prereplicative complex of yeast. |
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