首页> 外国专利> Identification and characterization of a novel human protein designated integrase interactor 1, that binds specifically to the human immunodeficiency virus type 1 integrase

Identification and characterization of a novel human protein designated integrase interactor 1, that binds specifically to the human immunodeficiency virus type 1 integrase

机译:鉴定和表征新型人类蛋白,称为整合酶相互作用子1,该蛋白与人免疫缺陷病毒1型整合酶特异性结合

摘要

Upon entry into a host cell, retroviruses direct the reverse transcription of the viral RNA genome and the establishment of an integrated proviral DNA. The retroviral integrase protein (IN) is responsible for the insertion of the viral DNA into host chromosomal targets. The IN catalyzes two specific biochemical reactions: (i) cleavage of the 3'termini of the viral DNA to produce 3'-OH ends, and (ii) joining of the two newly generated 3'-termini to the 5'-phosphates on each strand of the target sequence in a concerted strand-transfer reaction. The yeast two-hybrid system was used to identify a novel human gene product, herein designated integrase interactor 1 or INI-1, that binds tightly to the human immunodeficiency virus type 1 (HIV-1) integrase in vitro. Approximately 10.sup.6 complementary DNAs (cDNAs) of the HL60 macrophage-monocytic cell line were expressed as GAL4AC (activation domain) fusions and tested for coactivation of a reporter gene together with a GAL4DB (DNA binding) IN fusion. Overlapping cDNA clones were identified and their nucleotide sequences ascertained. Nucleotide sequence analysis revealed that INI-1 displays limited amino acid homology to the yeast SNF5 protein, a transcriptional activator required for high-level expression of many disparate cellular genes. The INI-1 gene product will prove useful for the generation of biochemical reagents and the development novel HIV-1 antiviral agents.
机译:进入宿主细胞后,逆转录病毒将指导病毒RNA基因组的逆转录并建立整合的前病毒DNA。逆转录病毒整合酶蛋白(IN)负责将病毒DNA插入宿主染色体靶标。 IN催化两个特定的生化反应:(i)裂解病毒DNA的3'末端以产生3'-OH末端,和(ii)将两个新生成的3'-末端连接到5'-磷酸在协同链转移反应中靶序列的每条链。酵母双杂交系统用于鉴定新的人类基因产物,在本文中称为整合酶相互作用子1或INI-1,其在体外与人免疫缺陷病毒1型(HIV-1)整合酶紧密结合。 HL60巨噬细胞-单核细胞系的大约10.sup.6个互补DNA(cDNA)被表达为GAL4AC(激活域)融合蛋白,并测试了报告基因与GAL4DB(DNA结合)IN融合蛋白的共激活。鉴定出重叠的cDNA克隆并确定其核苷酸序列。核苷酸序列分析表明,INI-1与酵母SNF5蛋白之间存在有限的氨基酸同源性,酵母SNF5蛋白是许多不同细胞基因高水平表达所需的转录激活因子。 INI-1基因产物将被证明对生化试剂的产生和新型HIV-1抗病毒剂的开发有用。

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