Sequence requirement for replication initiation at the rat aldolase B locus implicated in its functional correlation with transcriptional regulation.
Transcription promoter of the aldolase B gene was previously shown to be centered on an initiation region of DNA replication in rat hepatoma cells in vivo. Here, we defined an essential region required for replication in a plasmid form upon transfection. Deletion analyses around the origin region revealed that the proximal 200 bp promoter was necessary, but not sufficient for replication as flanking sequence restored replication activity. Therefore, the 200 bp region seemed to cooperate with the flanking sequence to play an important role in replication. Electrophoretic mobility shift assays using nuclear extracts from synchronously growing hepatoma cells showed that some protein factors bound to this region in a cell cycle-regulated manner. Since transcription of the aldolase B gene is repressed in the hepatoma cells, the cell cycle-regulated protein-binding is considered to be involved in regulation of replication initiation.
Zhao
Y
Y
Institute for Cell Biology and Genetics, Faculty of Agriculture, Iwate University, Japan.
Miyagi
S
S
Kikawada
T
T
Tsutsumi
K
K
D000818:Animals; D019556:COS Cells; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000378:metabolism; D002455:Cell Division; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism; D005634:Fructose-Bisphosphate Aldolase / Q000096:biosynthesis* / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D008114:Liver Neoplasms, Experimental; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014162:Transfection
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D005634:Fructose-Bisphosphate Aldolase