长的5'前导序列会抑制哺乳动物tRNA 3'加工内切核糖核酸酶从前体tRNA上去除3'拖尾序列。
Long 5' leaders inhibit removal of a 3' trailer from a precursor tRNA by mammalian tRNA 3' processing endoribonuclease.
作者信息
Nashimoto M, Wesemann D R, Geary S, Tamura M, Kaspar R L
机构信息
Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
出版信息
Nucleic Acids Res. 1999 Jul 1;27(13):2770-6. doi: 10.1093/nar/27.13.2770.
Mammalian tRNA 3' processing endoribonuclease (3' tRNase) can remove a 3' trailer from various pre-tRNAs without 5' leader nucleotides. To examine how 5[prime] leader sequences affect 3' processing efficiency, we performed in vitro 3' processing reactions with purified pig 3' tRNase and pre-tRNAArgs containing a 13-nt 3' trailer and a 5[prime] leader of various lengths. The 3' processing was slightly stimulated by 5[prime] leaders containing up to 7 nt, whereas leaders of 9 nt or longer severely inhibited the reaction. Structure probing indicated that the 5' leader sequences had little effect on pre-tRNA folding. Similar results were obtained using pre-tRNA(Val)s containing a 5' leader of various lengths. We also investigated whether 3'tRNase can remove 3' trailers that are stably base-paired with 5' leaders to form an extended acceptor stem. Even such small 5' leaders as 3 and 6 nt, when base-paired with a 3' trailer, severely hindered removal of the 3' trailer by 3' tRNase.
哺乳动物tRNA 3'加工内切核糖核酸酶（3' tRNase）可以从各种不含5'前导核苷酸的前体tRNA中去除3'拖尾序列。为了研究5'前导序列如何影响3'加工效率，我们使用纯化的猪3' tRNase和含有13个核苷酸的3'拖尾序列以及不同长度5'前导序列的前体tRNAArg进行了体外3'加工反应。含有多达7个核苷酸的5'前导序列对3'加工有轻微的促进作用，而9个核苷酸或更长的前导序列则严重抑制该反应。结构探测表明，5'前导序列对前体tRNA折叠影响很小。使用含有不同长度5'前导序列的前体tRNAVal也得到了类似的结果。我们还研究了3'tRNase是否能够去除与5'前导序列稳定碱基配对形成延伸受体茎的3'拖尾序列。即使是像3和6个核苷酸这样短的5'前导序列，当与3'拖尾序列碱基配对时，也会严重阻碍3' tRNase去除3'拖尾序列。
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