SPLICED LEADER RNA SEQUENCES CAN SUBSTITUTE FOR THE ESSENTIAL 5' END OF U1 RNA DURING SPLICING IN A MAMMALIAN INVITRO SYSTEM
SPLICED LEADER RNA SEQUENCES CAN SUBSTITUTE FOR THE ESSENTIAL 5' END OF U1 RNA DURING SPLICING IN A MAMMALIAN INVITRO SYSTEM
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DOI：
10.1016/0092-8674(90)90264-f
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发表时间：
1990-09-07
期刊：
影响因子：
64.5
通讯作者：
STEITZ, JA
中科院分区：
文献类型：
--
作者：
BRUZIK, JP;STEITZ, JA
在HeLa细胞核提取物中，与腺病毒内含子和3外显子连接的Leptomonascollosoma或Caelophabditiselegans SL RNA序列被高效且准确地剪接。使用RNAase H和与U1的前12个核苷酸互补的脱氧寡核苷酸灭活U1 snRNP后，含SL RNA的构建体的剪接继续不减少，而对照底物不再剪接。由于使用抗（U1）RNP抗体既未检测到U1 snRNP的结合也未检测到剪接的抑制，因此含有SL RNA的构建体的剪接可能完全不依赖于U1 snRNP。对变异L. collosoma构建体揭示了5“剪接位点周围的序列不足以赋予U1非依赖性剪接;迄今为止鉴定的最小的U1非依赖性区域仅保留SL RNA的第一个茎环。负责识别5“剪接位点的序列可以在剪接底物本身内重新定位，这加强了II组自我剪接和剪接体介导的前mRNA剪接之间的相似性。
Leptomonas collosoma or Caenorhabditis elegans SL RNA sequences joined to an adenovirus intron and 3 exon are spliced highly efficiently and accurately in HeLa nuclear extract. After inactivation of U1 snRNPs using RNAase H and a deoxyoligonucleotide complementary to the first 12 nucleotides of U1, splicing of SL RNA-containing constructs continues undiminished, whereas control substrates no longer splice. Since neither binding of U1 snRNPs nor inhibition of splicing is detected using anti-(U1)RNP antibodies, splicing of SL RNA-containing constructs may be entirely U1 snRNP independent. Analyses of altered L. collosoma constructs revealed that the sequence surrounding the 5'' splice site is not sufficient to confer U1-independent splicing; the smallest U1-independent region identified so far retains only the first stem-loop of the SL RNA. That sequences responsible for recognition of the 5'' splice site can be relocated within the splicing substrate itself reinforces the similarity between group II self-splicing and spliceosome-mediated pre-mRNA splicing.