Cloning and expression of the rat liver cDNA for peroxisomal enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase in lambda GT11. Transcriptional regulation of enzyme activity by Wy-14643 in primary cultures of rat hepatocytes.
Cloning and expression of the rat liver cDNA for peroxisomal enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase in lambda GT11. Transcriptional regulation of enzyme activity by Wy-14643 in primary cultures of rat hepatocytes.
复制标题
大鼠肝脏过氧化物酶体烯酰辅酶 A 水合酶、3-羟酰辅酶 A 脱氢酶 cDNA 在 lambda GT11 中的克隆和表达。
DOI：
--
复制
发表时间：
1987
期刊：
影响因子：
--
通讯作者：
A. Roy
中科院分区：
文献类型：
--
作者：
B. Chatterjee;C. Murty;M. J. Olson;A. Roy
降血脂药物Wy-14643对大鼠肝脏过氧化物酶体的增殖与参与脂肪酸β -氧化的过氧化物酶体酶的伴随诱导有关。为了探究这一诱导过程的分子机制，我们在lambda gt11表达载体上克隆了过氧化物酶体双功能酶烯酰辅酶a水合酶、3-羟基酰基辅酶a脱氢酶（ECH）的cDNA。用兔抗ECH单特异性血清筛选文库。杂交选择mrna翻译证实免疫反应克隆含有ECH双功能酶的cDNA序列。克隆的cDNA用于确定大鼠肝细胞原代培养中与酶诱导相关的早期事件。用ECH cDNA探针对肝细胞总RNA进行斑点杂交显示，在与Wy-14643孵育约10-15 h后，ECH mRNA开始升高。孵育24 h和48 h时，与对照相比，ECH mRNA的刺激分别达到26倍和47倍。肝细胞离体核的径流实验显示，药物治疗后5小时，ECH基因的转录率没有增加，在药物治疗后10小时和20小时，ECH基因的转录率分别增加了2倍和11倍。从这些结果我们得出结论，Wy-14643增加ECH活性是由于ECH基因转录率的提高。然而，肝细胞暴露于Wy-14643后相对较长的滞后期（约10-15小时）表明，药物对ECH mRNA的诱导可能涉及间接影响该基因的转录。
Proliferation of rat liver peroxisomes by the hypolipidemic drug Wy-14643 is associated with a concomitant induction of peroxisomal enzymes involved in the beta-oxidation of fatty acids. In order to explore the molecular mechanism of this induction process we have cloned the cDNA for the peroxisomal bifunctional enzyme enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase (ECH) in the lambda gt11 expression vector. The library was screened with the monospecific rabbit antiserum to ECH. Hybrid-selected-mRNA translation established that the immunoreactive clones contain the cDNA sequences of the ECH bifunctional enzyme. The cloned cDNA was used to define the early events associated with enzyme induction in primary cultures of rat hepatocytes. Dot-blot hybridization of the total hepatocyte RNA with the ECH cDNA probe showed that the ECH mRNA begins to rise at about 10-15 h following incubation with Wy-14643. At 24 h and 48 h of incubation the stimulation of the ECH mRNA over the vehicle-treated control reached 26-fold and 47-fold respectively. Run-off experiments in the isolated nuclei of hepatocytes showed no increase in the transcription rate of the ECH gene at 5 h after drug treatment and a 2-fold and 11-fold increase at 10 h and 20 h of drug treatment. From these results we conclude that the increase in ECH activity by Wy-14643 is due to an enhancement of the rate of transcription of the ECH gene. However, the relatively long lag period of about 10-15 h after exposure of hepatocytes to Wy-14643 suggests that the induction of the ECH mRNA may involve an indirect effect of the drug on the transcription of this gene.
登录
查看更多内容
降血脂药物 Wy-14,643 诱导过氧化物酶体和非过氧化物酶体蛋白的肝脏信使 RNA 种类发生可逆改变。
DOI：
10.1042/bj2140879
发表时间：
1983
期刊：
The Biochemical journal
影响因子：
--
作者：
Chatterjee,B;Demyan,WF;Lalwani,ND;Reddy,JK;Roy,AK
通讯作者：
Roy,AK
DOI：
10.1073/pnas.83.6.1747
发表时间：
1986-03-01
影响因子：
11.1
作者：
REDDY, JK;GOEL, SK;RAO, MS
通讯作者：
RAO, MS
雄激素抑制性大鼠肝脏蛋白 SMP-2 的 cDNA 分子克隆和表征。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Chatterjee,B;Majumdar,D;Ozbilen,O;Murty,CV;Roy,AK
通讯作者：
Roy,AK
细胞病理学中的过氧化物酶体（微生物）。
DOI：
--
发表时间：
1984
期刊：
International review of experimental pathology
影响因子：
--
作者：
Goldfischer,S;Reddy,JK
通讯作者：
Reddy,JK
DOI：
10.1073/pnas.80.5.1194
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
YOUNG, RA;DAVIS, RW
通讯作者：
DAVIS, RW