Wu Yiqin, Fa Ming, Tae Eunju Lee, Schultz Peter G, Romesberg Floyd E
Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA.
J Am Chem Soc. 2002 Dec 11;124(49):14626-30. doi: 10.1021/ja028050m.
In an effort to expand the genetic alphabet, a number of unnatural, predominantly hydrophobic, nucleoside analogues have been developed which pair selectively in duplex DNA and during enzymatic synthesis. Significant progress has been made toward the efficient in vitro replication of DNA containing these base pairs. However, the in vivo expansion of the genetic alphabet will require that the unnatural nucleoside triphosphates be available within the cell at sufficient concentrations for DNA replication. We report our initial efforts toward the development of an unnatural in vivo nucleoside phosphorylation pathway that is based on nucleoside salvage enzymes. The first step of this pathway is catalyzed by the D. melanogaster nucleoside kinase, which catalyzes the phosphorylation of nucleosides to the corresponding monophosphates. We demonstrate that each unnatural nucleoside is phosphorylated with a rate that should be sufficient for the in vivo replication of DNA.
为了扩展遗传字母表，人们已经开发出了多种非天然的、主要为疏水性的核苷类似物，它们在双链DNA中以及酶促合成过程中能够选择性配对。在含有这些碱基对的DNA的高效体外复制方面已经取得了重大进展。然而，遗传字母表在体内的扩展将需要细胞内有足够浓度的非天然核苷三磷酸用于DNA复制。我们报告了我们在基于核苷补救酶开发非天然体内核苷磷酸化途径方面的初步努力。该途径的第一步由黑腹果蝇核苷激酶催化，它将核苷磷酸化为相应的单磷酸。我们证明，每种非天然核苷的磷酸化速率应足以用于DNA的体内复制。
