Evtushenko V I, Khanson K P
Biokhimiia. 1988 May;53(5):862-71.
Using [14C]adenine as a labeled precursor, the biosynthesis and processing of heterogeneous nuclear RNA (hnRNA) in chromatin and nucleoplasm were studied. It was shown that intraperitoneally injected actinomycin D (50-500 micrograms/100 g of body weight) did not inhibit the biosynthesis of hnRNA and rRNA in rat thymocytes. Besides biosynthesis, the polyadenylation of the bulk of primary transcripts was also localized in the chromatin. rRNA precursors constituted only a small part of the newly synthesized hnRNA molecules. During processing, highly polymeric (28S-55S) poly(A+)-hnRNA fractions passed from chromatin to the nucleoplasm, as a result of which a large number of high molecular weight polyadenylated hnRNA molecular were pooled in the nucleoplasm. Thymocyte hnRNA was tightly bound to nuclear structures and its isolation tributed stringent conditions. The bulk of poly(A+)-hnRNA molecules were degraded without formation of stable intermediates. The pattern of poly(A-)-hnRNA maturation in chromatin and nucleoplasm was identical, i.e., in both compartments the decrease in the size and accumulation of processed 16S-20S RNA molecules was observed. About 6% of newly synthesized poly(A+)-hnRNA was converted into polysomal mRNA. Cytoplasmic poly(A+)-RNA consists of two fractions, i.e., metabolically stable high molecular weight (greater than or equal to 40S) RNA molecules and a less stable 10S-20S RNA.
以[14C]腺嘌呤作为标记前体,研究了染色质和核质中不均一核RNA(hnRNA)的生物合成及加工过程。结果表明,腹腔注射放线菌素D(50 - 500微克/100克体重)并不抑制大鼠胸腺细胞中hnRNA和rRNA的生物合成。除了生物合成外,大部分初级转录本的聚腺苷酸化也定位于染色质中。rRNA前体仅占新合成的hnRNA分子的一小部分。在加工过程中,高度聚合的(28S - 55S)聚(A+)-hnRNA组分从染色质转移到核质,结果大量高分子量的聚腺苷酸化hnRNA分子聚集在核质中。胸腺细胞hnRNA与核结构紧密结合,其分离需要严格的条件。大部分聚(A+)-hnRNA分子在没有形成稳定中间体的情况下被降解。染色质和核质中聚(A-)-hnRNA的成熟模式是相同的,即在这两个区室中都观察到加工后的16S - 20S RNA分子大小的减小和积累。新合成的聚(A+)-hnRNA中约6%转化为多聚核糖体mRNA。细胞质聚(A+)-RNA由两个组分组成,即代谢稳定的高分子量(大于或等于40S)RNA分子和较不稳定的10S - 20S RNA。