De Greve J, Battey J, Fedorko J, Birrer M, Evan G, Kaye F, Sausville E, Minna J
National Cancer Institute and Naval Hospital, Bethesda, Maryland 20814.
Mol Cell Biol. 1988 Oct;8(10):4381-8. doi: 10.1128/mcb.8.10.4381-4388.1988.
The human proto-oncogene L-myc generates at least four different mRNAs by alternative RNA processing. We have identified two phosphorylated L-myc proteins with molecular masses of 60,000 and 66,000 daltons [p60L-myc(human) and p66L-myc(human)] in a small-cell carcinoma line expressing high levels of L-myc mRNA. These proteins have a short half-life and are localized to the nuclear matrix fraction, as previously reported for the c-myc and N-myc proteins. In vitro translation experiments demonstrated that both the p60 and p66 species are encoded by a 3.9-kilobase (kb) mRNA which retains intron 1, while only the p60 protein is translated from a 3.6-kb L-myc mRNA which has had intron 1 removed. While L-myc proteins [p32L-myc(human) and p37L-myc(human)] could be synthesized in vitro from 2.2-kb mRNA templates, no such proteins were detected by immunoprecipitation in vivo. These observations suggest that alternative RNA processing of the L-myc transcript could play a role in determining the steady-state levels of the p60L-myc and p66L-myc proteins.
人类原癌基因L-myc通过可变RNA加工产生至少四种不同的mRNA。我们在一个表达高水平L-myc mRNA的小细胞癌系中鉴定出两种磷酸化的L-myc蛋白,分子量分别为60,000和66,000道尔顿[p60L-myc(人类)和p66L-myc(人类)]。这些蛋白半衰期短,定位于核基质部分,正如先前报道的c-myc和N-myc蛋白一样。体外翻译实验表明,p60和p66蛋白均由保留内含子1的3.9千碱基(kb) mRNA编码,而只有p60蛋白是从去除了内含子1的3.6 kb L-myc mRNA翻译而来。虽然L-myc蛋白[p32L-myc(人类)和p37L-myc(人类)]可以在体外从2.2 kb mRNA模板合成,但在体内免疫沉淀未检测到此类蛋白。这些观察结果表明,L-myc转录本的可变RNA加工可能在决定p60L-myc和p66L-myc蛋白的稳态水平中起作用。