Hiley Shawna L, Babak Tomas, Hughes Timothy R
Banting and Best Department of Medical Research, University of Toronto 112 College Street, Toronto, ON M5G 1L6, Canada.
Nucleic Acids Res. 2005 May 26;33(9):3048-56. doi: 10.1093/nar/gki608. Print 2005.
We used a microarray containing probes that tile all known yeast noncoding RNAs (ncRNAs) to investigate RNA biogenesis on a global scale. The microarray verified a general loss of Box C/D snoRNAs in the TetO7-BCD1 mutant, which had previously been shown for only a handful of snoRNAs. We also monitored the accumulation of improperly processed flank sequences of pre-RNAs in strains depleted for known RNA nucleases, including RNase III, Dbr1p, Xrn1p, Rat1p and components of the exosome and RNase P complexes. Among the hundreds of aberrant RNA processing events detected, two novel substrates of Rnt1p (the RUF1 and RUF3 snoRNAs) were identified. We also identified a relationship between tRNA 5' end processing and tRNA splicing, processes that were previously thought to be independent. This analysis demonstrates the applicability of microarray technology to the study of global analysis of ncRNA synthesis and provides an extensive directory of processing events mediated by yeast ncRNA processing enzymes.
我们使用了一个包含能覆盖所有已知酵母非编码RNA(ncRNA)的探针的微阵列,以在全球范围内研究RNA生物合成。该微阵列证实了TetO7-BCD1突变体中Box C/D snoRNA普遍缺失,此前仅在少数几个snoRNA中观察到这种情况。我们还监测了在缺失已知RNA核酸酶(包括RNase III、Dbr1p、Xrn1p、Rat1p以及外切体和RNase P复合物的组分)的菌株中,前体RNA加工不当的侧翼序列的积累情况。在检测到的数百个异常RNA加工事件中,鉴定出了Rnt1p的两个新底物(RUF1和RUF3 snoRNA)。我们还确定了tRNA 5'端加工与tRNA剪接之间的关系,这两个过程以前被认为是相互独立的。该分析证明了微阵列技术在ncRNA合成全局分析研究中的适用性,并提供了由酵母ncRNA加工酶介导的加工事件的广泛目录。