Nucleic Acids Research, Vol 24, Issue 10 1887-1894, Copyright © 1996 by Oxford University Press
SE Waltz, AA Trivedi and M Leffak
The origin of replication of the c-myc gene in HeLa cells was previously
identified at low resolution within 3.5 kb 5' to the P1 promoter, based on
replication fork polarity and the location of DNA nascent strands. To
define the initiation events in the c-myc origin at higher resolution the
template bias of nascent DNAs in a 12 kb c-myc domain has been analyzed by
hybridization to strand specific probes. Strong switches in the asymmetry
of nascent strand template preference confirm that replication initiates
non-randomly at multiple sites within 2.4 kb 5' to the c-myc P1 promoter,
and at other sites over a region of 12 kb or more. The strongest template
biases occur in the 2.4 kb region 5' of the P1 promoter, shown earlier to
contain sequences which allow the autonomous semiconservative replication
of c-myc plasmids. An asymmetric pyrimidine heptanucleotide consensus
sequence has been identified which occurs 12 times in the c-myc origin
zone, and whose polarity exactly correlates with the polarity of nascent
strand synthesis.
ARTICLES
DNA replication initiates non-randomly at multiple sites near the c-myc gene in HeLa cells
Department of Biochemistry and Molecular Biology, Wright State University, Dayton, OH 45435, USA.
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