Sburlati A R, Manrow R E, Berger S L
Section on Genes and Gene Products, National Cancer Institute, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):253-7. doi: 10.1073/pnas.88.1.253.
The function of prothymosin alpha has been investigated by using four different antisense oligodeoxyribonucleotides directed at selected regions of its mRNA. In every case, when synchronized human myeloma cells were released from stationary phase by incubation in fresh medium containing antisense oligomers, cell division was prevented or inhibited; sense oligomers and random antisense oligomers had no effect. A detailed analysis of synchronized cell populations indicated that sense-treated and untreated cells divided approximately 17 hr after growth initiation, whereas cells incubated with antisense oligomer 183, a 16-mer targeted 5 bases downstream of the initiation codon, entered mitosis approximately one cell division late. The failure to divide correlated directly with a deficit in prothymosin alpha and with the continued presence of intact intracellular antisense oligomers over a period of at least 24 hr. Because antisense oligomers had no effect either on the timing of the induction of prothymosin alpha mRNA upon growth stimulation or on mRNA levels seen throughout the cell cycle, we concluded that antisense DNA caused specific hybrid arrest of translation. Our data suggest that prothymosin alpha is required for cell division. However, there is no evidence that prothymosin alpha directly regulates mitosis.
通过使用针对其mRNA选定区域的四种不同反义寡脱氧核糖核苷酸，对原胸腺素α的功能进行了研究。在每种情况下，当同步化的人骨髓瘤细胞在含有反义寡聚物的新鲜培养基中孵育而从静止期释放时，细胞分裂被阻止或抑制；正义寡聚物和随机反义寡聚物则没有作用。对同步化细胞群体的详细分析表明，经正义处理和未处理的细胞在生长开始后约17小时分裂，而与反义寡聚物183（一种针对起始密码子下游5个碱基的16聚体）孵育的细胞进入有丝分裂大约晚一个细胞周期。无法分裂与原胸腺素α的缺乏以及完整的细胞内反义寡聚物在至少24小时内的持续存在直接相关。由于反义寡聚物对生长刺激后原胸腺素α mRNA诱导的时间或整个细胞周期中所见的mRNA水平均无影响，我们得出结论，反义DNA导致了特异性的翻译杂交阻滞。我们的数据表明原胸腺素α是细胞分裂所必需的。然而，没有证据表明原胸腺素α直接调节有丝分裂。
