Sinnett, D., Woolf, E., Xie, W., Glatt, K., Kirkness, E. F., Nielsen, T. O., Zannis-Hadjopoulos, M., Price, G. B., Lalande, M.
Identification of a Putative DNA Replication Origin In the Gamma-aminobutyric Acid Receptor Subunit Beta3 and Alpha5 Gene Cluster on Human chromosome 15q11-q13, a Region Associated With Parental Imprinting and Allele-specific Replication Timing
Gene. 1996 Sep 16; 173(2): 171-7.
The region containing the GABAA receptor beta3 and alpha5 subunit-encoding genes is subject to parental imprinting and is organized in different allele-specific replication timing domains. A 60-kb domain displaying a maternal early/paternal late pattern of allele-specific replication timing asynchrony is nested within a larger region displaying the opposite pattern. The proximal portion of this maternal early replicating domain is incorporated into phage clone lambda84. In order to identify DNA structures which may be associated with the boundary between the replication domains, phage lambda84 has been subcloned into smaller fragments and several of these have been analyzed by nucleotide sequencing. A plot of helical stability for 13kb of contiguous sequence reveals several A + T-rich regions which display potential DNA unwinding. The plasmid subclones from phage lambda84 have been analyzed for bent DNA and one of these, p82, contains bent DNA and overlaps with the region of highest potential helical instability. Of the seven plasmids tested, only p82 shows strong autonomous replication activity in an in vitro replication assay, with replication initiating within the genomic insert. These results suggest that a putative origin of DNA replication contained within p82 may play a role in establishing the allele-specific replication timing domains in the GABAA receptor subunit gene cluster.