Lusis A J, Chapman V M, Herbstman C, Paigen K
J Biol Chem. 1980 Oct 10;255(19):8959-62.
Genetic studies have shown that the induction of beta-glucuronidase in mouse kidney in response to androgens is under the control of the Gus-r gene, closely linked to the beta-glucuronidase structural gene, Gus-s, on mouse Chromosome 5. Despite the fact that a single structural gene codes for beta-glucuronidase, enzyme molecules show considerable charge heterogeneity, presumably as a result of post-translational events. Because the enzyme is a tetramer, this heterogeneity has complicated examination of the cis versus trans action of the Gus-r gene. In order to resolve the problem, we have developed a quantitative method for determining the amount of beta-glucuronidase derived from each structural allele in mice heterozygous for beta-glucuronidase electrophoretic mobility. The method involves separation and quantitation of genetically variant cyanogen bromide peptides from the enzyme. Using this technique, we show that Gus-r acts cis, suggesting that it directly affects the template activity of the glucuronidase structural gene.
遗传学研究表明，小鼠肾脏中β-葡萄糖醛酸酶因雄激素诱导的表达受Gus-r基因控制，该基因与小鼠5号染色体上的β-葡萄糖醛酸酶结构基因Gus-s紧密连锁。尽管单个结构基因编码β-葡萄糖醛酸酶，但酶分子表现出相当大的电荷异质性，这可能是翻译后事件的结果。由于该酶是四聚体，这种异质性使对Gus-r基因顺式与反式作用的研究变得复杂。为了解决这个问题，我们开发了一种定量方法，用于确定β-葡萄糖醛酸酶电泳迁移率杂合的小鼠中每个结构等位基因衍生的β-葡萄糖醛酸酶量。该方法包括从酶中分离和定量遗传变异的溴化氰肽。使用这项技术，我们表明Gus-r基因起顺式作用，这表明它直接影响葡萄糖醛酸酶结构基因的模板活性。
