MAMMALIAN MITOCHONDRIAL TRANSFER-RNAS - CHROMATOGRAPHIC PROPERTIES, SIZE AND ORIGIN
MAMMALIAN MITOCHONDRIAL TRANSFER-RNAS - CHROMATOGRAPHIC PROPERTIES, SIZE AND ORIGIN
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DOI：
10.1093/nar/6.2.455
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发表时间：
1979-01-01
影响因子：
14.9
通讯作者：
FREEMAN, KB
中科院分区：
文献类型：
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作者：
AUJAME, L;FREEMAN, KB
离体大鼠肝线粒体与放射性氨基酸孵育导致精氨酸、天冬酰胺、亮氨酸、赖氨酸、甲硫氨酸、脯氨酸和缬氨酸的tRNA充电。通过RPC-5上的色谱法显示氨酰-tRNA与其胞质对应物不同。通过尿素聚丙烯酰胺平板凝胶电泳，发现所有这些线粒体氨酰-tRNA的长度约为70-76个核苷酸。先前在哺乳动物细胞中未发现的独特线粒体天冬酰胺酰-和脯氨酰-tRNA已被证明与线粒体DNA杂交。线粒体亮氨酰-tRNA在RPC-5上分离成3个峰，第一种物质的分子大小和部分RNase T1消化模式与其他两种物质的组合不同。杂交的部分加和性表明，每一个都由mtDNA上的一个单独的基因编码。通过RPC-5层析分离的线粒体tRNAmMet和tRNAfMet的单独基因也被证明。这些结果使哺乳动物mtDNA编码的单个tRNA的数量达到21个。
Incubation of isolated rat liver mitochondria with radioactive amino acids resulted in the charging of tRNAs for arginine, asparagine, leucine, lysine, methionine, proline and valine. The aminoacyl-tRNAs were shown to be distinct from their cytosolic counterparts by chromatography on RPC-5. By electrophoresis on urea polyacrylamide slab gels it was found that all these mitochondrial aminoacyl-tRNAs were about 70–76 nucleotides long. The unique mitochondrial asparaginyl- and prolyl-tRNAs, not previously identified in mammalian cells, were shown to hybridize to mtDNA. Mitochondrial leucyl-tRNA separated into 3 peaks on RPC-5 and the first species was shown to be different than a combination of the other two by molecular size and partial RNase T1digestion patterns. Each was coded by a separate gene on mtDNA as shown by partial additivity of hybridization. Separate genes for mitochondrial tRNAmMetand tRNAfMetseparated by RPC-5 chromatography, were also demonstrated. These results bring to 21 the number of individual tRNAs coded by mammalian mtDNA.