NEGATIVE-STRAND RNA TRANSCRIPTS ARE PRODUCED IN HUMAN-IMMUNODEFICIENCY-VIRUS TYPE 1-INFECTED CELLS AND PATIENTS BY A NOVEL PROMOTER DOWN-REGULATED BY TAT
NEGATIVE-STRAND RNA TRANSCRIPTS ARE PRODUCED IN HUMAN-IMMUNODEFICIENCY-VIRUS TYPE 1-INFECTED CELLS AND PATIENTS BY A NOVEL PROMOTER DOWN-REGULATED BY TAT
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DOI：
10.1128/jvi.68.2.979-987.1994
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发表时间：
1994-02-01
影响因子：
5.4
通讯作者：
REDFIELD, RR
中科院分区：
文献类型：
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作者：
MICHAEL, NL;VAHEY, MT;REDFIELD, RR
目前对人类免疫缺陷病毒1型（HIV-1）转录的理解是基于5'长末端重复序列的正链极性转录本的单向表达。我们现在报告的HIV-1转录本的负链极性获得从急性和慢性感染的细胞系，通过使用模板定向特异性逆转录酶-PCR检测。这些发现通过分析来自15名HIV-1感染患者的外周血单核细胞样品的RNA在自然感染中得到证实。从急性感染的A3.01细胞中分离出一个cDNA，它来自一个负链极性的2.3-kb聚腺苷酸化的HIV-I RNA，编码一个高度保守的189-氨基酸开放阅读框，与包膜基因反向平行。通过使用报告基因构建，我们进一步发现，一个新的负链启动子功能内的负响应元件的3'长末端重复，这是下调共表达的达特。定点突变实验证明NF-κ B I和USF位点对负链启动子活性至关重要。这些数据扩展了HIV-1的编码能力，并提出了一种反义调节病毒生命周期的科尔，
Current understanding of human immunodeficiency virus type 1 (HIV-I) transcription is based on unidirectional expression of transcripts with positive-strand polarity from the 5' long terminal repeat. We now report HIV-1 transcripts' with negative-strand polarity obtained from acutely and chronically infected cell lines by use of a template orientation-specific reverse transcriptase-PCR assay. These findings were confirmed in natural infection by analysis of RNA derived from peripheral blood mononuclear cell samples from 15 HIV-l-infected patients. A cDNA derived from a 2.3-kb polyadenylated HIV-I RNA with negative-strand polarity which encodes a highly conserved 189-amino-acid open reading frame antiparallel to the envelope gene was isolated from acutely infected A3.01 cells. Through use of reporter gene constructions, we further found that a novel negative-strand promoter functions within the negative response element of the 3' long terminal repeat, which is downregulated by coexpression of Tat. Site-directed mutagenesis experiments demonstrated that NF-kappa B I and USF sites are crucial for negative-strand promoter activity. These data extend the coding capacity of HIV-1 and suggest a cole for antisense regulation of the viral life cycle,