title: Drosophila follicle cell amplicons as models for metazoan DNA replication: a cyclinE mutant exhibits increased replication fork elongation.
abstract: Gene clusters amplified in the ovarian follicle cells of Drosophila serve as powerful models for metazoan DNA replication. In response to developmental signals, specific genomic regions undergo amplification by repeated firing of replication origins and bidirectional movement of replication forks for approximately 50 kb in each direction. Previous work focused on initiation of amplification, defining replication origins, establishing the role of the prereplication complex and origin recognition complex (ORC), and uncovering regulatory functions for the Myb, E2F1, and Rb transcription factors. Here, we exploit follicle cell amplification to investigate the control of DNA replication fork progression and termination, poorly understood processes in metazoans. We identified a mutant in which, during gene amplification, the replication forks move twice as far from the origin compared with wild type. This phenotype is the result of an amino acid substitution mutation in the cyclinE gene, cyclinE(1f36). The rate of oogenesis is normal in cyclinE(1f36)/cyclinE(Pz8) mutant ovaries, indicating that increased replication fork progression is due to increased replication fork speed, possibly from increased processivity. The increased amplification domains observed in the mutant imply that there are not replication fork barriers preventing replication forks from progressing beyond the normal 100-kb amplified region. These results reveal a previously unrecognized role for CyclinE in controlling replication fork movement.
authors.affiliation: Whitehead Institute and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.
authors.forename: Eugenia A
authors.initials: EA
authors.lastname: Park
authors.forename: David M
authors.initials: DM
authors.lastname: Macalpine
authors.forename: Terry L
authors.initials: TL
authors.lastname: Orr-Weaver
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D019927:Cyclin E / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis; D004261:DNA Replication; D004330:Drosophila / Q000166:cytology / Q000235:genetics*; D029721:Drosophila Proteins / Q000737:chemistry / Q000378:metabolism; D005260:Female; D018628:Gene Dosage; D008957:Models, Genetic; D008969:Molecular Sequence Data; D005810:Multigene Family; D009154:Mutation / Q000235:genetics*; D006080:Ovarian Follicle / Q000166:cytology* / Q000378:metabolism*; D010063:Ovum / Q000166:cytology / Q000254:growth & development; D010641:Phenotype; D021381:Protein Transport
chemical_list: D019927:Cyclin E; D029721:Drosophila Proteins; D004247:DNA; D001973:Bromodeoxyuridine