An overlapping set of DNA elements in the rat aldolase B gene origin/promoter regulates transcription and autonomous replication.
Promoter of the rat aldolase B (AldB) gene is centered on an origin of chromosomal DNA replication in vivo, and it directs autonomous replication upon transfection into cultured cells. Previous studies showed that the 200 bp promoter fragment is necessarily required for the autonomous replication. Here, we identified three cis-elements required for replication within the 200 bp promoter, using autonomously replicating plasmids carrying various mutations and deletions. One is an element that is previously defined as a regulatory element for liver-specific transcription (site C). Other two, purine-rich (site PPu) and A (T)-rich (site A/T) sequences, were those often found in eukaryotic origin regions. Sites C and PPu were found to bind specific nuclear factors in transfected cells, and the results of competitive binding assay implied direct or indirect interaction between sites C and PPu.
Miyagi
S
S
Cryobiosystem Research Center, Faculty of Agriculture, Ueda, 020-8550, Japan.
Zhao
Y
Y
Saitoh
Y
Y
Tsutsumi
K
K
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D019556:COS Cells; D002522:Chlorocebus aethiops; D004261:DNA Replication; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014162:Transfection
D011994:Recombinant Proteins; D005634:Fructose-Bisphosphate Aldolase