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关岛患者大脑细胞质颗粒组分中的RNA指导的DNA聚合酶活性。

RNA-instructed DNA polymerase activity in a cytoplasmic particulate fraction in brains from Guamanian patients.

作者信息

Viola M V, Frazier M, White L, Brody J, Spiegelman S

出版信息

J Exp Med. 1975 Aug 1;142(2):483-94. doi: 10.1084/jem.142.2.483.

Abstract

Nervous system tissues from a number of patients with idiopathic neurological disorders were examined for biochemical evidence of RNA tumor virus infection. RNase-sensitive DNA polymerase activity was found in a cytoplasmic particulate fraction from two patients with Guamanian amyotrophic lateral sclerosis (ALS) but not in brains from two normal U.S. individuals. The buoyant density of the enzyme-containing fraction was 1.16-1.18 g/ml and could be converted to a denser region of the gradient (1.24 g/ml) by treatment with the nonionic surfactant, Sterox. The cation and detergent requirements for the endogenous RNase-sensitive DNA polymerase reaction were determined. The early (5 min) endogenous reverse transcriptase product was analyzed by cesium sulfate gradient centrifugation. RNase- and heat-sensitive RNA-DNA hybrids were detected in the product analysis of two ALS, one Parkinsonism-dementia (PD) brain, and two brains from asymptomatic Chamorros but not in brains from normal U.S. individuals and a number of patients with neuro-psychiatric disorders. The DNA product was a 4.5S heteropolymer that hybridized more extensively to RNA extracted from the enzyme-containing pellet from PD brain as compared to a similar fraction from normal U.S. brain. The DNA product appeared to be unrelated to Rausvher or visna virus 70S RNA as determined by RNA-[-3H]DNA hybridization.

摘要

对一些患有特发性神经疾病患者的神经系统组织进行检查,以寻找RNA肿瘤病毒感染的生化证据。在两名关岛肌萎缩侧索硬化症(ALS)患者的细胞质颗粒部分发现了对核糖核酸酶敏感的DNA聚合酶活性,但在两名美国正常人的大脑中未发现。含酶部分的浮力密度为1.16 - 1.18克/毫升,通过用非离子表面活性剂Sterox处理可将其转化为梯度中密度更高的区域(1.24克/毫升)。确定了内源性对核糖核酸酶敏感的DNA聚合酶反应对阳离子和去污剂的需求。通过硫酸铯梯度离心分析早期(5分钟)内源性逆转录酶产物。在两名ALS患者、一名帕金森病痴呆(PD)患者的大脑以及两名无症状查莫罗人的大脑产物分析中检测到了对核糖核酸酶和热敏感的RNA - DNA杂交体,但在美国正常人的大脑以及一些患有神经精神疾病患者的大脑中未检测到。DNA产物是一种4.5S杂聚物,与从PD大脑含酶沉淀中提取的RNA相比,与从美国正常大脑类似部分提取的RNA杂交更广泛。通过RNA - [ - 3H]DNA杂交确定,该DNA产物似乎与劳斯肉瘤病毒或维斯纳病毒70S RNA无关。

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