Establishment of high sensitive Salmonella tester strains expressing mammalian acetvltransferase and sulfotransferase

表达哺乳动物乙酰转移酶和磺基转移酶的高灵敏沙门氏菌测试菌株的建立

• 批准号: 05557120
• 负责人: KATO Ryuichi
• 金额: $ 9.86万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05557120/
• 关键词: Ames test; Mutagenicity; Carcinogenicity; Drug metabolizing enzymes; Metabolic activation; Acetyltransferase; Sulfotransferase; Gene expression; Anes試験; アセチル化酵素; アセチル化酵素遺伝子; 異所遺伝子発現; アリルアミン; N,O‐アセチル転移反応; サルモネラ菌; ベンチジン

We have been studying metabolic activation of carcinogenic arylamines in mammalian livers and Salmonella. We reported that N,O-acetyltransferase and sulfotransferase, which playd a role in the metabolic activation in mammalian livers, were deficient in Ames tester strains. In this project, we newly established two Ames tester strains in which hamster arylamine acetyltransferase and rat N-hydroxy arylamine O-sulfotransferase were independently expressed. The new strains were 100-10,000times more sensitive to environmental carcinogens including N-hydroxy-2-acetylaminofluorene. Furthermore, the acetyltransferase-expressing strain showed high sensitivity to benzidine to which no Ames tester strain had showed any sensitivity.The acetyltransferase and the sulfotransferase were shown to catalyze activation of N-hydroxy derivatives of not only arylamine (amide) s but also food-derived pyrolysate carcinogens.These results indicated that the expression of mammalian esterification enzymes in Salmonella Ames tester strain resulted in new highly sensitive tester strains toward various kinds of environmental carcinogens. Thus, we concluded that this kind of approach is useful for the more predictive assessment of environmental mutagens and carcinogens.

我们一直在研究哺乳动物肝和沙门氏菌中致癌芳胺的代谢激活。我们报道说，在哺乳动物肝脏的代谢激活中扮演的N，O-乙酰基转移酶和磺基转移酶在AMES tester菌株中不足。在这个项目中，我们新建立了两个AMES测试仪菌株，其中仓鼠芳胺乙酰转移酶和大鼠N-羟基芳基芳基胺O-磺胺转移酶被独立表达。新菌株对包括N-羟基-2-乙酰氨基氟的环境致癌物更敏感。此外，表达乙酰基转移酶的菌株对苯甲胺表现出很高的敏感性，没有AMES测试剂菌株显示出任何敏感性。乙酰基转移酶和磺胺转移酶被证明可以催化催化N-羟基衍生物的激活，不仅是食物（胺）S的激活。这些结果表明，沙门氏菌AMES测试仪菌株中哺乳动物酯化酶的表达导致新的高度敏感的测试仪菌株对各种环境致癌物。因此，我们得出的结论是，这种方法对于对环境诱变剂和致癌物的更具预测性评估很有用。

项目成果

Yamazoe,Y.: "Proceedings of the 23rd International Symposium of the Princess Takamatsu Cancer Research Fund" Princeton Scientific Publishing Co., Inc. (in press),

Yamazoe,Y.：“第 23 届高松公主癌症研究基金国际研讨会论文集”普林斯顿科学出版有限公司（正在印刷中），

Ozawa, S.: "Activation of 2-hydroxyamino-1-methyl-6-phenylimidazo [4,5-b] pyridine by cDNA-expressed human and rat arylsulfotransferases." Jpn.J.Cancer Res.85. 1220-1228 (1994)

Ozawa, S.：“cDNA 表达的人和大鼠芳基磺基转移酶激活 2-羟基氨基-1-甲基-6-苯基咪唑[4,5-b]吡啶。”

Yamazoe, Y.: "Structural similarity and diversity of sulfotransferases." Chem.-Biol.Interact.92. 107-117 (1994)

Yamazoe, Y.：“磺基转移酶的结构相似性和多样性。”

Ozawa,S.: "Activation of 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine by cDNA-expressed human and rat arylsulfotransferases." Jpn. J. Cancer Res.85. 1220-1228 (1994)

Ozawa,S.：“cDNA 表达的人和大鼠芳基磺基转移酶激活 2-羟基氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶。”

Yamazoe, Y.: "Sulfotransferase- and acetyltransferase-mediated activation of carcinogenic N-hydroxyarylamines in mammals and bacteria, and their modulation by thiols. Antimutagenesis and Anticarcinogenesis Mechanisms III" Plenum Press. 161-169 (1993)

Yamazoe, Y.：“哺乳动物和细菌中致癌 N-羟芳胺的磺基转移酶和乙酰基转移酶介导的激活，及其硫醇的调节。抗突变和抗癌机制 III”Plenum Press。