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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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Information & Authors

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Published In

Science
Volume 287 | Issue 5460
17 March 2000

Submission history

Received: 6 October 1999
Accepted: 2 February 2000
Published in print: 17 March 2000

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Authors

Affiliations

Gulnara Abdurashidova*
Molecular Biology and
Marta Deganuto*
Molecular Biology and
Raffaella Klima
Molecular Biology and
Silvano Riva
Istituto di Genetica Biochimica ed Evoluzionistica of the Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy.
Giuseppe Biamonti
Istituto di Genetica Biochimica ed Evoluzionistica of the Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy.
Mauro Giacca
Molecular Medicine Units, International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34012 Trieste, Italy.
Arturo Falaschi
Molecular Biology and

Notes

*
These authors contributed equally to this work.
To whom correspondence should be addressed. E-mail: [email protected]

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