Assembly of functionally active Drosophila origin recognition complex from recombinant proteins.
In eukaryotes the sites for the initiation of chromosomal DNA replication are believed to be determined in part by the binding of a heteromeric origin recognition complex (ORC) to DNA. We have cloned the genes encoding the subunits of the Drosophila ORC. Each of the genes is unique and can be mapped to discrete chromosomal locations implying that the pattern and developmental regulation of origin usage in Drosophila is not regulated solely by a large family of different ORC proteins. The six-subunit ORC can be reconstituted with recombinant proteins into a complex that restores DNA replication in ORC-depleted Drosophila or Xenopus egg extracts.
Chesnokov
I
I
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California 94720 USA.
Gossen
M
M
Remus
D
D
Botchan
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004261:DNA Replication / Q000502:physiology; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics*; D004330:Drosophila / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D051738:Origin Recognition Complex; D010063:Ovum / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013094:Spermatozoa / Q000378:metabolism; D013997:Time Factors; D014981:Xenopus / Q000378:metabolism
D004268:DNA-Binding Proteins; D051738:Origin Recognition Complex; D011993:Recombinant Fusion Proteins