Millette R L, Klaiber R
J Virol. 1980 Jun;34(3):604-14. doi: 10.1128/JVI.34.3.604-614.1980.
The transcriptional organization of the genome of herpes simplex virus type 1 was analyzed by measuring the sensitivity of viral polypeptide synthesis to UV irradiation of the infecting virus. Herpes simplex virus type 1 was irradiated with various doses of UV light and used to infect xeroderma pigmentosum fibroblasts. Immediate early transcription units were analyzed by having cycloheximide present throughout the period of infection, removing the drug at 8 h postinfection, and pulse-labeling proteins with [35S]methionine. Delayed early transcription units were analyzed in similar studies by having 9-beta-D-arabinofuranosyladenine present during the experiment to block replication of the input irradiated genome. The viral polypeptides were separated by gel electrophoresis and quantitated by densitometry of the gel autoradiograms. The following results were obtained. (i) The UV target sizes for the viral transcription units analyzed ranged from 1.44 to 5.65 kilobase pairs. This implies that the corresponding primary transcripts have minimum molecular weights ranging from 0.46 x 10(6). (ii) The genes for the four viral proteins, 165, 145, 116, and 71 (molecular weight x 10(3), exhibited UV target sizes that agree with their calculated gene size or measured mRNA size or both and thus must reside in promoter-adjacent positions. (iii) The transcription units for the remaining genes analyzed showed target sizes that range from 0.42 to 2.59 kilobase pairs greater than needed to encode the respective proteins. This probably is a reflection of their distances from promoters or the presence of intervening sequences or both. It further suggests that these genes are transcribed as precursor RNA molecules that are larger than their mRNA's. (iv) The results indicate that none of the immediate early genes analyzed can be cotranscribed, whereas some of the delayed early genes might be cotranscribed. No evidence was found for the existance of large, multigene transcription units.
通过测量病毒多肽合成对感染病毒紫外线照射的敏感性，对1型单纯疱疹病毒基因组的转录组织进行了分析。用不同剂量的紫外线照射1型单纯疱疹病毒，并用于感染着色性干皮病成纤维细胞。通过在整个感染期间存在环己酰亚胺、在感染后8小时去除该药物并用[35S]甲硫氨酸脉冲标记蛋白质来分析立即早期转录单位。在类似研究中，通过在实验期间存在9-β-D-阿拉伯呋喃糖基腺嘌呤以阻断输入的受照射基因组的复制来分析延迟早期转录单位。通过凝胶电泳分离病毒多肽，并通过凝胶放射自显影片的光密度测定进行定量。获得了以下结果。（i）所分析的病毒转录单位的紫外线靶标大小范围为1.44至5.65千碱基对。这意味着相应的初级转录本的最小分子量范围为0.46×10(6)。（ii）四种病毒蛋白165、145、116和71（分子量×10(3)）的基因表现出与它们计算的基因大小或测量的mRNA大小或两者一致的紫外线靶标大小，因此必须位于启动子相邻位置。（iii）所分析的其余基因的转录单位显示靶标大小比编码各自蛋白质所需的大小大0.42至2.59千碱基对。这可能反映了它们与启动子的距离或间隔序列的存在或两者。这进一步表明这些基因转录为比它们的mRNA更大的前体RNA分子。（iv）结果表明，所分析的立即早期基因中没有一个可以共转录，而一些延迟早期基因可能是共转录的。没有发现存在大的多基因转录单位的证据。
