Cook W J, Coen D M
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, 02115, USA.
Virology. 1996 Apr 1;218(1):204-13. doi: 10.1006/viro.1996.0180.
Using Northern blot, primer extension, and S1 nuclease analyses of wild-type and deletion-containing herpes simplex type 1 viruses, we found that UL24 sequences are contained in six different transcripts that originate from three previously identified mRNA start sites. Thus, the six UL24 transcripts represent three different pairs of 5' coterminal mRNAs. Each transcript pair consists of a short species whose 3' end corresponds to a polyadenylation signal located just downstream of the UL24 open reading frame, and a longer species whose 3' end corresponds to a polyadenylation signal located downstream of the UL26 gene. Maximal accumulation of the short UL24 transcripts was at early times during infection, while accumulation of the longer species did not decrease at late times. Consistent with early kinetics, the short transcripts were less sensitive to drugs that inhibited viral DNA replication than the longer transcripts which exhibited leaky-late kinetics. Quantitative S1 nuclease analysis indicated that 3' ends corresponding to the UL24 polyadenylation site were significantly more abundant at early times than at late times. Thus, differential polyadenylation determines the kinetics of accumulation of different UL24 transcripts.
通过对野生型和含缺失的1型单纯疱疹病毒进行Northern印迹、引物延伸和S1核酸酶分析，我们发现UL24序列存在于六种不同的转录本中，这些转录本源自三个先前确定的mRNA起始位点。因此，这六种UL24转录本代表了三对不同的5' 共末端mRNA。每对转录本由一个短转录本和一个长转录本组成，短转录本的3' 末端对应于位于UL24开放阅读框下游的一个聚腺苷酸化信号，长转录本的3' 末端对应于位于UL26基因下游的一个聚腺苷酸化信号。短UL24转录本在感染早期积累达到最大值，而长转录本在后期积累并未减少。与早期动力学一致，短转录本对抑制病毒DNA复制的药物的敏感性低于表现出渗漏晚期动力学的长转录本。定量S1核酸酶分析表明，对应于UL24聚腺苷酸化位点的3' 末端在早期比在后期明显更丰富。因此，差异聚腺苷酸化决定了不同UL24转录本积累的动力学。
