POLYCYCLIC HYDROCARBON METABOLISM AND CARCINOGENESIS

该实验室的总体研究目标是了解 细胞色素P450 IA 1是一种单加氧酶， 在活化多环烃，如苯并（a） 芘（BP）。 我们假设，一个胞质4S多环 碳氢化合物结合蛋白作为P45 OIA 1的反式激活因子发挥作用 基因 本提案的具体目标与这一假设有关。 的 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， 这种蛋白质，并确定是否磷酸化和/或与 hsp 90可以改变其活性; B）确定5 '-上游 与4s蛋白结合的P450 IA 1基因的区域，即，多环 碳氢化合物响应元件（PRE），并通过体外核 转录测定表明，4s蛋白是这种转录的反式激活因子。 c）从肝cDNA和基因组文库中分离4s蛋白基因 并表征该基因，和d）确定转染的影响， 能够转录4s基因的反义基因并导入的载体 4s蛋白的抗体对P450 IA 1基因的诱导作用， 对BP治疗的反应。 这些研究应确立一个核心作用， 对于4s多环烃结合蛋白在调节 P450 IA 1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.