Lin J, Vogt V M
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853, USA.
Mol Cell Biol. 1998 Oct;18(10):5809-17. doi: 10.1128/MCB.18.10.5809.
PpLSU3, a mobile group I intron in the rRNA genes of Physarum polycephalum, also can home into yeast chromosomal ribosomal DNA (rDNA) (D. E. Muscarella and V. M. Vogt, Mol. Cell. Biol. 13:1023-1033, 1993). By integrating PpLSU3 into the rDNA copies of a yeast strain temperature sensitive for RNA polymerase I, we have shown that the I-PpoI homing endonuclease encoded by PpLSU3 is expressed from an RNA polymerase I transcript. We have also developed a method to integrate mutant forms of PpLSU3 as well as the Tetrahymena intron TtLSU1 into rDNA, by expressing I-PpoI in trans. Analysis of I-PpoI expression levels in these mutants, along with subcellular fractionation of intron RNA, strongly suggests that the full-length excised intron RNA, but not RNAs that are further cleaved, serves as or gives rise to the mRNA.
多头绒泡菌rRNA基因中的一个可移动的I类内含子PpLSU3,也能归巢到酵母染色体核糖体DNA(rDNA)中(D. E. 马斯卡雷拉和V. M. 沃格特,《分子与细胞生物学》13:1023 - 1033,1993年)。通过将PpLSU3整合到对RNA聚合酶I温度敏感的酵母菌株的rDNA拷贝中,我们已经表明,PpLSU3编码的I - PpoI归巢内切酶是从RNA聚合酶I转录本表达的。我们还开发了一种方法,通过反式表达I - PpoI,将PpLSU3的突变形式以及四膜虫内含子TtLSU1整合到rDNA中。对这些突变体中I - PpoI表达水平的分析,以及内含子RNA的亚细胞分级分离,强烈表明全长切除的内含子RNA,而不是进一步切割的RNA,作为mRNA或产生mRNA。