Padgett R A, Podar M, Boulanger S C, Perlman P S
Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas 75235.
Science. 1994 Dec 9;266(5191):1685-8. doi: 10.1126/science.7527587.
The stereochemical specificities and reaction courses for both self-splicing steps of a group II intron have been determined by phosphorothioate substitution at the 5' and 3' splice site phosphodiester bonds. Both steps of the splicing reaction proceeded with a phosphorothioate in the Sp configuration but were blocked by the Rp diastereomer. Both steps also proceeded with inversion of stereochemical configuration around phosphorus, consistent with a concerted transesterification reaction. These results are identical to those found for nuclear precursor mRNA (pre-mRNA) splicing and provide support for the hypothesis that group II introns and nuclear pre-mRNA introns share a common evolutionary history.
通过对II组内含子自身剪接步骤中5'和3'剪接位点磷酸二酯键进行硫代磷酸酯取代,确定了这两个剪接步骤的立体化学特异性和反应过程。剪接反应的两个步骤在Sp构型的硫代磷酸酯存在下都能进行,但被Rp非对映体阻断。两个步骤在磷周围的立体化学构型也发生了反转,这与协同转酯反应一致。这些结果与核前体mRNA(前体mRNA)剪接的结果相同,并为II组内含子和核前体mRNA内含子具有共同进化历史的假说提供了支持。