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首页> 外文期刊>Molecular and Cellular Biology >Characterization of a fusion cDNA (RARA/myl) transcribed from the t(15;17) translocation breakpoint in acute promyelocytic leukemia.
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Characterization of a fusion cDNA (RARA/myl) transcribed from the t(15;17) translocation breakpoint in acute promyelocytic leukemia.

机译：从急性早幼粒细胞白血病的t（15; 17）易位断点转录的融合cDNA（RARA / myl）的表征。

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A nonrandom chromosomal translocation breakpoint, t(15;17)(q22;q21), is found in almost all patients with acute promyelocytic leukemia (APL). Most of these breakpoints occur within the second intron of the retinoic acid receptor-alpha (RARA) gene. We screened a cDNA library of APL and have identified and sequenced a cDNA transcribed from the t(15;17) translocation breakpoint. The 5' end of cDNA p1715 consists of 503 bp of the RARA exon II sequence. A 1.76-kb cDNA without homology to any known gene available in GenBank was found truncated downstream. This cDNA sequence was assigned to chromosome 15 by dot blot hybridization of the flow cytometry-sorted chromosomes. We designate this fusion cDNA RARA/myl, which is different from myl/RARA reported by de The et al. (H. de The, C. Chomienne, M. Lanotte, L. Degos, and A. Dejean, Nature (London) 347:558-561, 1990). This result demonstrates that the two different types of hybrid mRNA can be transcribed from this breakpoint. We screened a non-APL cDNA library and identified a 2.8-kb myl cDNA. This cDNA is able to encode a polypeptide with a molecular weight of 78,450. Alternative splicing of the myl gene which resulted in myl proteins with different C terminals was found. Southern blot analysis of the genomic DNA isolated from 17 APL patients by using the myl DNA probe demonstrated that the myl gene in 12 samples was rearranged. Northern (RNA) blot analysis of RARA gene expression in two APL RNA samples showed abnormal mRNA species of 4.2 and 3.2 kb in one patient and of 4.8 and 3.8 kb in another patient; these were in addition to the normal mRNA species of 3.7 and 2.7-kb. The myl DNA probe detected a 2.6-kb abnormal mRNA in addition to the normal mRNA species of 3.2, 4.2, and 5.5 kb. Using the polymerase chain reaction, we demonstrated that both RARA/myl and myl/RARA were coexpressed in samples from three different APL patients. From this study, we conclude that the t(15;17) translocation breakpoint results in the transcription of two different fusion transcripts which are expected to be translated into fusion proteins.
机译：在几乎所有急性早幼粒细胞白血病（APL）患者中都发现了一个非随机染色体易位断点t（15; 17）（q22; q21）。这些断点大多数发生在视黄酸受体α（RARA）基因的第二个内含子内。我们筛选了APL的cDNA文库，并鉴定并测序了从t（15; 17）易位断点转录的cDNA。 cDNA p1715的5'端由503 bp的RARA外显子II序列组成。发现一个1.76kb cDNA与GenBank中可用的任何已知基因没有同源性，在下游被截断。通过流式细胞仪分类的染色体的斑点印迹杂交将该cDNA序列分配给15号染色体。我们指定了该融合cDNA RARA / myl，它与de The et al。报道的myl / RARA不同。 （H.de The，C.Chomienne，M.Lanotte，L.Degos和A.Dejean，Nature（伦敦）347：558-561，1990）。该结果表明可以从该断点转录两种不同类型的杂交mRNA。我们筛选了非APL cDNA文库，并鉴定了2.8-kb myl cDNA。该cDNA能够编码分子量为78450的多肽。发现了myl基因的选择性剪接，其导致具有不同C末端的myl蛋白。使用myl DNA探针对17例APL患者的基因组DNA进行Southern印迹分析，结果表明12个样品中的myl基因被重新排列。在两个APL RNA样品中进行RARA基因表达的Northern（RNA）印迹分析显示，一名患者的异常mRNA分别为4.2和3.2 kb，另一名患者的4.8和3.8 kb。这些是3.7和2.7-kb正常mRNA的补充。除了正常的3.2、4.2和5.5 kb的mRNA种类以外，myl DNA探针还检测到2.6 kb的异常mRNA。使用聚合酶链反应，我们证明了RARA / myl和myl / RARA在三个不同APL患者的样品中共表达。从这项研究中，我们得出的结论是，t（15; 17）易位转折点导致两种不同融合转录本的转录，这些转录本有望被翻译成融合蛋白。

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