HTLV-III SEQUENCES AT THE TRANSCRIPTIONAL LEVEL

机会性病毒感染在慢性阻塞性肺疾病患者中很普遍 获得性免疫缺陷综合征(艾滋病)。为了调查 疱疹病毒组的机会性感染的可能性 可能诱导人类T淋巴细胞病毒的表达 我们构建了III型(HTLV-III)长末端重复序列(LTR) 含有HTLV-III的永久性细胞系(小鼠和猿猴) 氯霉素乙酰转移酶(CAT)基因的启动子。 而在瞬时基因表达分析中，HTLV-III LTR 表达CAT活性与猴病毒-40(SV40)一样高 启动子，染色质后未观察到CAT活性 在永久细胞系中整合。这些潜伏期的重叠感染 含有疱疹病毒株的HTLV-III LTR重新激活 用S1核酸酶RNA分析确定HTLV-III的LTR值。 鉴于猴病毒-40(SV40)和腺病毒感染 含潜伏HTLV-III LTR的细胞株不能激活HTLV- III LTR的表达。潜伏的HTLV-III LTR的再激活 5氮杂胞苷、放线菌亚胺后也观察到转录 和紫外线照射处理。在分离的环境中运行转录 细胞核，表明激活是在转录水平上。 单纯疱疹病毒1型(HSV-1)需要病毒激活 病毒决定的即刻早期(IE)基因表达 放线菌亚胺存在时的感染和突变体 病毒基因表达有缺陷。该计划的长远目标 这项建议中描述的研究是为了阐明 激活HTLV-III LTR的机制(S) 转录水平。具体目标是确定 与潜伏的HTLV-III LTR有关的病毒基因产物 转录激活；建立永久细胞系 含有HTLV-III LTR的人B细胞和T细胞；鉴定 抑制HTLV-III调节区的机制(S)和 确定涉及抑制的HTLV-III LTRDNA序列 和重新激活。初步结果表明， 疱疹病毒组的机会性感染可能会导致 潜伏感染HTLV的个体中HTLV-III的表达 表格，并指出这些细胞系在研究 其他理化刺激对HTLV-III再激活的影响。

Opportunistic viral infections are prevalent in patients with acquired immune deficiency syndrome (AIDS). To investigate the possibility that opportunistic infections in the herpesvirus group might induce expression from the human T-lymphotropic virus type III (HTLV-III) long-terminal repeats (LTR), we constructed permanent cell lines (murine and simian), containing the HTLV-III LTR directing the chloramphenicol acetyltransferase (CAT) gene. Whereas in transient gene expression assays, the HTLV-III LTR express CAT activity as high as the simian virus-40 (SV40) early promoter, no CAT activity is observed after chromatin integration in permanent cell lines. Superinfection of these latent HTLV-III LTR containing lines with herpes viruses reactivated the HTLV-III LTR as determined by S1 nuclease RNA analysis. Whereas simian virus-40 (SV40) and adenovirus infection of the latent HTLV-III LTR containing cell lines did not activate HTLV- III LTR expression. Reactivation of latent HTLV-III LTR transcription is also observed after 5 azacytidine, cycloheximide and UV irradiation treatments. Run-on transcription in isolated nuclei, suggests that the activation is on the transcriptional level. Activation by herpes simplex virus type 1 (HSV-1) required viral immediate early (IE) gene expression as determined by virus infection in the presence of cycloheximide and with mutants defective in viral gene expression. The long-term objective of the research described in this proposal is to elucidate the mechanism(s) by which the HTLV-III LTR could be activated on the transcriptional level. The specific aims are to identify the viral gene products responsible for latent HTLV-III LTR transcription activation; to establish permanent cell lines in human B and T-cells containing the HTLV-III LTR; to identify the mechanism(s) of repression of the HTLV-III regulatory region and to define the HTLV-III LTR DNA sequences involved repression and reactivation. The preliminary results suggest that opportunistic infection of the herpesvirus group could induce HTLV-III expression in individuals harboring the virus in a latent form and points to the potential of these cell lines to study the affects of other physiochemical stimuli on HTLV-III reactivation.