Light activation of transcription: photocaging of nucleotides for control over RNA polymerization.
We describe the use of ATP caged with [7-(diethylamino)coumarin-4-yl]methyl (DEACM) for light-controlled in vitro transcription reactions. Polymerization is blocked when DEACM is bonded to the gamma phosphate group of the ATP molecule. Controlled light irradiation releases ATP and transcription is initiated. In order to provide full control over the process, conditions involved in substrate release, nucleotide availability after release and the effect of the released coumarin in RNA polymerization were assessed in further detail. Together, our data provide the first direct evidence of control over enzymatic polymerization of nucleic acids through light. This approach may provide researchers with a unique tool for the study of biological processes at a molecular level.
Pinheiro
André Vidal
AV
REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa ,Campus de Caparica, 2829-516 Caparica, Portugal.
Baptista
Pedro
P
Lima
João Carlos
JC
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D003374:Coumarins / Q000737:chemistry*; D008027:Light; D010777:Photochemistry; D012313:RNA / Q000096:biosynthesis*; D014158:Transcription, Genetic / Q000528:radiation effects*; D015533:Transcriptional Activation
D003374:Coumarins; C531214:adenosine triphosphate (7-(diethylamino)coumarin-4-yl)methyl ester; D012313:RNA; D000255:Adenosine Triphosphate