Stavnezer J
Nucleic Acids Res. 1986 Aug 11;14(15):6129-44. doi: 10.1093/nar/14.15.6129.
RNA blotting was employed to examine polyadenylated immunoglobulin alpha chain RNAs in a B lymphoma synthesizing membrane-bound and secretory IgA and in a hybridoma which synthesizes predominantly secretory IgA. Both cell lines were derived from the I.29 lymphoma and expressed the identical heavy chain variable region gene. In addition to the predicted mRNA precursors, four novel species of polyadenylated alpha RNAs were detected. The presence of a RNA species which was too large to have the same 3' end as the largest mRNA for membrane-bound alpha chain (alpha m) implied that transcription continued past the alpha m poly(A) site, and that such transcripts could be polyadenylated. Alternatively, transcription of this alpha RNA was initiated 5' to the normal cap site. Two species of RNA were detected which encoded the alpha m domain and the intervening sequence between the alpha constant (C alpha) and alpha m domain but not the C alpha domain. These RNA molecules were of sizes appropriate for their derivation by endonucleolytic cleavage of a precursor for alpha m mRNA at the poly(A) site of the mRNA for secreted alpha chains (alpha s). The presence of these three alpha RNA species suggested that alternative and successive cleavage/polyadenylation events could occur on a single transcript to produce either alpha m or alpha s mRNAs. An additional novel species of RNA was detected which indicated that the order of removal of the large IVSs did not always proceed in the 5' to 3' direction.
采用RNA印迹法检测了一个合成膜结合型和分泌型IgA的B淋巴瘤细胞系以及一个主要合成分泌型IgA的杂交瘤细胞系中多聚腺苷酸化免疫球蛋白α链RNA。这两个细胞系均源自I.29淋巴瘤,且表达相同的重链可变区基因。除了预测的mRNA前体,还检测到了四种新的多聚腺苷酸化αRNA。存在一种RNA分子,其大小过大,以至于其3'末端与膜结合型α链(αm)的最大mRNA的3'末端不同,这意味着转录在αm多聚腺苷酸化位点之后仍在继续,并且这样的转录本可以被多聚腺苷酸化。或者,这种αRNA的转录是在正常帽位点的5'端起始的。检测到两种RNA,它们编码αm结构域以及α恒定区(Cα)和αm结构域之间的间隔序列,但不编码Cα结构域。这些RNA分子的大小适合于通过在分泌型α链(αs)mRNA的多聚腺苷酸化位点对αm mRNA前体进行核酸内切酶切割而产生。这三种αRNA的存在表明,在单个转录本上可能发生交替和连续的切割/多聚腺苷酸化事件,以产生αm或αs mRNA。还检测到另一种新的RNA,这表明大内含子序列(IVS)的去除顺序并不总是从5'端向3'端进行。