CO2 response for expression of ribulose-1,5-bisphosphate carboxylase/oxygenase genes is inhibited by AT-rich decoy in the cyanobacterium.
The CO(2)-regulatory function of the AT-rich element in the promoter for ribulose-1,5-bisphosphate carboxylase/oxygenase (rbc) genes in the cyanobacterium Synechococcus sp. PCC7002 was analyzed using the transcription factor decoy approach. Double-stranded phosphorothioate AT-rich oligonucleotides with high affinity for a sequence-specific DNA-binding protein were successfully introduced into cyanobacterial cells in culture without any transfection reagent. The AT-rich decoy oligonucleotides interfered with CO(2) regulation of rbc expression by blocking the binding of the sequence-specific DNA-binding protein, indicating that the AT-rich element plays a critical role in CO(2) regulation for rbc genes. The decoy oligonucleotide approach to cyanobacteria provides a simple and excellent tool for investigating transcriptional regulation in vivo.
Biological Science Laboratories, Toray Research Center, Inc., 1111 Tebiro, Kamakura, Kanagawa 248-8555, Japan. takuo_onizuka@trc.toray.co.jp
Takuo
T
Onizuka
Hideo
H
Akiyama
Sumiyo
S
Endo
Shozo
S
Kanai
Masahiko
M
Hirano
Satoshi
S
Tanaka
Hitoshi
H
Miyasaka
D020856:AT Rich Sequence; D001692:Biological Transport; D002245:Carbon Dioxide / Q000037:antagonists & inhibitors*; D000458:Cyanobacteria / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D004794:Enzyme Repression; D016650:Fluorescein-5-isothiocyanate; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D017930:Genes, Reporter; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D020218:Response Elements; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics*; D015533:Transcriptional Activation / Q000187:drug effects
D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D002245:Carbon Dioxide; D012273:Ribulose-Bisphosphate Carboxylase; D016650:Fluorescein-5-isothiocyanate