Start sites of bidirectional DNA synthesis at the human lamin B2 origin.
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.
Abdurashidova
G
G
Molecular Biology and Molecular Medicine Units, International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34012 Trieste, Italy.
Deganuto
M
M
Klima
R
R
Riva
S
S
Biamonti
G
G
Giacca
M
M
Falaschi
A
A
D020856:AT Rich Sequence; D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis; D017931:DNA Primers / Q000378:metabolism; D004261:DNA Replication; D004640:Emetine / Q000494:pharmacology; D016193:G1 Phase; D006367:HeLa Cells; D006801:Humans; D034921:Lamin Type B; D034882:Lamins; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D011485:Protein Binding; D018741:Replication Origin; D016196:S Phase
D017931:DNA Primers; D034921:Lamin Type B; D034882:Lamins; D009687:Nuclear Proteins; C022262:Okazaki fragments; D004247:DNA; D004640:Emetine