POLYCYCLIC HYDROCARBON METABOLISM AND CARCINOGENESIS

本实验室的总体研究目标是了解 细胞色素P450IA1的调节机制 在多环烃活化中的作用，如苯并(A) 芘(BP)。我们推测胞液中的4S多环 烃结合蛋白作为P45OIA1的反式激活剂 吉恩。这项提议的具体目标与这一假设有关。这个 具体目的是：a)确定4S的氨基酸序列 多环烃结合蛋白，产生多克隆抗体 这一蛋白质，并确定是否磷酸化和/或结合 HSP90可以改变其活性；b)确定5‘上游的性质 P450IA1基因与4S蛋白结合的区域，即多环 碳氢化合物反应元件(Pre‘s)，并通过体外核建立 转录分析表明，4S蛋白是这一过程的反式激活因子 C)从肝脏cDNA库和基因组文库中分离4S蛋白基因 并对该基因进行特征分析，以及d)确定转染组的效果 转录反义4S基因的载体和导入 抗4S蛋白抗体对小鼠P450IA1基因诱导的影响 对BP治疗的反应。这些研究应确立核心作用 4S多环烃结合蛋白在细胞周期调控中的作用 P450IA1基因的表达。

The overall research objective of this laboratory is to understand the mechanism of regulation of cytochrome P450IA1, a monooxygenase that functions in the activation of polycyclic hydrocarbons, such as benzo (a) pyrene (BP). We have postulated that a cytosolic 4S polycyclic hydrocarbon-binding protein functions as a trans-activator of the P45OIA1 gene. The specific aims of this proposal relate to this hypothesis. The specific aims are to: a) determine the amino acid sequence of the 4S polycyclic hydrocarbon-binding protein, produce polyclonal antibodies to this protein, and determine if phosphorylation and/or conjunction with hsp90 can modify its activity; b) ascertain the nature of the 5'-upstream regions of the P450IA1 gene to which the 4s protein binds, i.e., polycyclic hydrocarbon responsive elements (PRE's), and establish by in vitro nuclear transcriptional assays that the 4s protein is a trans-activator of this gene; c) isolate the 4s protein gene from liver cDNA and genomic libraries and characterize this gene, and d) determine the effects of transfection of vectors that would transcribe antisense to the 4s gene and of introduction of antibodies to the 4s protein on the induction of the P450IA1 gene in response to BP treatment. These studies should establish a central role for the 4s polycyclic hydrocarbon-binding protein in the regulation of the expression of the P450IA1 gene.