METHYLATION OF NUCLEAR SIMIAN VIRUS-40 RNAS
METHYLATION OF NUCLEAR SIMIAN VIRUS-40 RNAS
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DOI：
10.1128/jvi.32.1.52-60.1979
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发表时间：
1979-01-01
影响因子：
5.4
通讯作者：
KHOURY, G
中科院分区：
文献类型：
--
作者：
ALONI, Y;DHAR, R;KHOURY, G
在最佳条件下用[甲基 - ³H]甲硫氨酸和[³H]尿苷进行脉冲标记的核RNA，是从感染猴空泡病毒40（SV - 40）后期的[非洲绿猴肾]细胞中制备的。标记的RNA与SV - 40 DNA的限制性片段进行杂交。将与特定限制性片段的早期（E）链或晚期（L）链退火的甲基 - ³H标记的RNA水平，与与同一片段退火的[³H]尿苷标记的RNA进行比较。与各种限制性片段杂交的甲基 - ³H标记的RNA被洗脱，并分析其帽结构和内部的6 - 甲基腺苷残基。L链和E链转录本的帽结构主要位于含有病毒DNA复制起点的一个片段中。SV - 40核RNA中内部6 - 甲基腺苷残基的最高水平位于83 - 0.0图距单位的一个片段内的L链上，以及位于37 - 67图距单位的一个片段内的E链的RNA转录本中。这两个区域的部分代表在细胞质mRNA中不存在的间隔序列。在其他位置发现较低水平的内部甲基化。讨论了内部6 - 甲基腺苷残基在RNA加工以及从细胞核到细胞质的转运中的潜在作用。
Nuclear RNAs, pulse-labeled with [methyl-3H]methionine and [3H]uridine under optimal conditions, were prepared from [African green monkey kidney] cells infected with SV-40 at a late time after infection. The labeled RNAs were hybridized with restriction fragments of SV-40 DNA. Levels of methyl-3H-labeled RNA annealing with the early (E) or late (L) strands of a particular restriction fragment were compared with [3H] uridine-labeled RNA annealing with the same fragment. The methyl-3H-labeled RNA hybridizing to the various restriction fragments was eluted and analyzed for caps and internal 6-methyladenosine residues. Caps of transcripts of the L and E strands were primarily located in a fragment containing the origin for viral DNA replication. The highest level of the internal 6-methyladenosine residues in SV-40 nuclear RNA was located on the L strand within a fragment from 83-0.0 map units and in RNA transcripts from the E strand within a fragment from 37-67 map units. Portions of both these regions represent intervening sequences not represented in cytoplasmic mRNA''s. Lower levels of internal methylation were found in other locations. The potential role of internal 6-methyladenosine residues in RNA processing and transport from the nucleus to the cytoplasm is discussed.