Citation:

Light activation of transcription: photocaging of nucleotides for control over RNA polymerization, Pinheiro, {André Vidal}, Baptistap Pedro, and Lima {João Carlos} , Nucleic Acids Research, aug, Volume 36, Number 14, (2008)

Abstract:

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.

Notes:

The authors would like to acknowledge Dr A. Jorge Parola for fruitful suggestions related to photochemistry procedures, Dr João Noronha for Mass Spectra analysis, Dr Isabel Sá Nogueira and Dr José Inácio for assistance with a-32P-UTP experiments. This work was supported by Fundac¸ ão para a Ciência e Tecnologia (REQUIMTE, CIGMH, PTDC/BIO/66514/2006, PTDC/SAU-BEB/ 66511/2006, FRH/BD/24276/2005 to A.V.P.); Fundac¸ ão Calouste Gulbenkian (Ref. 76436). Funding to pay the Open Access publication charges for this article was provided by Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa.