Grant D F, Hammock B D
Department of Environmental Toxicology, University of California, Davis 95616.
Mol Gen Genet. 1992 Aug;234(2):169-76. doi: 10.1007/BF00283836.
The amount of glutathione S-transferase-2 (GST-2) protein and enzyme activity in a mutant strain (strain GG) of the yellow fever mosquito (Aedes aegypti) is approximately 25-fold higher than in the wild-type (++) strain. The mode of inheritance of the GG phenotype was studied in F1 and backcross progeny using GST enzyme assays, isozyme-specific antisera, and Northern blot analysis. Enzyme assay of parental and F1 progeny showed that the ++ phenotype was dominant to the GG phenotype. This was true for larvae as well as for all tissues examined in adults in both sexes. Immunoblotting experiments showed that, like the ++ strain, F1 larvae and adults express very low levels of GST-2 protein compared with the GG strain. Northern blotting experiments showed that the steady-state levels of GST-2 mRNA in parental and F1 hybrid larvae closely matched the enzyme activity and immunological data. These results suggest the existence of a trans-acting regulatory locus that acts to repress GST-2 mRNA transcription and/or decrease GST-2 mRNA stability in ++ and F1 hybrids. GST enzyme activity in backcross progeny, however, did not segregate into the two distinct phenotypes (low and high) predicted for a single locus, dominant allele model. Backcross progeny expressed a wide range of GST activity and GST-2 protein amount with no apparent fit to simple Mendelian ratios. These backcross data suggest that additional loci are also involved in regulating GST-2 isozyme expression.(ABSTRACT TRUNCATED AT 250 WORDS)
黄热病蚊子(埃及伊蚊)的一个突变株(GG株)中谷胱甘肽S-转移酶-2(GST-2)蛋白的含量和酶活性比野生型(++株)高约25倍。使用GST酶测定、同工酶特异性抗血清和Northern印迹分析,在F1和回交后代中研究了GG表型的遗传模式。对亲本和F1后代的酶测定表明,++表型对GG表型呈显性。在幼虫以及两性成虫中检查的所有组织中都是如此。免疫印迹实验表明,与GG株相比,F1幼虫和成虫与++株一样,表达的GST-2蛋白水平非常低。Northern印迹实验表明,亲本和F1杂交幼虫中GST-2 mRNA的稳态水平与酶活性和免疫学数据密切匹配。这些结果表明存在一个反式作用调节位点,该位点在++和F1杂种中起抑制GST-2 mRNA转录和/或降低GST-2 mRNA稳定性的作用。然而,回交后代中的GST酶活性并没有分离成单基因座显性等位基因模型预测的两种不同表型(低和高)。回交后代表达了广泛的GST活性和GST-2蛋白量,与简单的孟德尔比率没有明显的契合度。这些回交数据表明,其他基因座也参与调节GST-2同工酶的表达。(摘要截短至250字)