Structural homologies and functional similarities between mammalian origins of replication and amplification promoting sequences.
MuNTS2, a 423 bp sequence isolated from the non-transcribed spacer of murine rDNA stimulates the amplification of cis-linked plasmid DNA in mouse cells under selective conditions. Here we demonstrate that a 180 bp subdomain of muNTS2 is highly homologous (approximately 70%) to three domains of the first well-characterized origin of replication of mammalian chromosomes, i.e. the origin of bidirectional replication (OBR) of the dihydrofolate reductase (DHFR) locus in Chinese hamster ovary (CHO) cells. When subcloned, the 180 bp homology region of muNTS2 was revealed to be essential for the amplification promoting activity of muNTS2. Fragments of the initiation zone of DNA replication from the DHFR locus of hamster cells containing the domains of homology to the mouse muNTS2 element proved also to promote DNA amplification. Thus, the screening system for amplification promoting elements turned out to detect an origin of bidirectional replication.
Stolzenburg
F
F
Institut für Biochemie, Julius-Maximilians-Universität Würzburg, Germany.
Gerwig
R
R
Dinkl
E
E
Grummt
F
F
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002874:Chromosome Mapping / Q000379:methods; D006224:Cricetinae; D004261:DNA Replication / Q000502:physiology*; D004275:DNA, Ribosomal / Q000235:genetics; D005784:Gene Amplification / Q000235:genetics*; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D018741:Replication Origin / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection
D004275:DNA, Ribosomal; D013762:Tetrahydrofolate Dehydrogenase