首页   按字顺浏览 期刊浏览 卷期浏览 Characterization of human polymorphic DNA repair methyltransferase
Characterization of human polymorphic DNA repair methyltransferase

 

作者: Ryo Inoue,   Masako Abe,   Yusaku Nakabeppu,   Mutsuo Sekiguchi,   Teruaki Mori,   Tomokazu Suzuki,  

 

期刊: Pharmacogenetics  (OVID Available online 2000)
卷期: Volume 10, issue 1  

页码: 59-66

 

ISSN:0960-314X

 

年代: 2000

 

出版商: OVID

 

关键词: O6-methylguanine-DNA methyltransferase;DNA repair;polymorphism;alkylating agent

 

数据来源: OVID

 

摘要:

The O6-methylguanine-DNA methyltransferase (MGMT) is a critical defence against alkylation-induced mutagenesis and carcinogenesis. More than a 20-fold interindividual difference in the MGMT activity is known to exist among human cultured fibroblasts. We previously reported three allelic variants of the humanMGMTgene, namely V1, V2, and V3. Both V1 and V2 carry amino acid substitutions, Leu84Phe and Trp65Cys, respectively, while V3 has a silent mutation. In order to reveal the pharmacogenetic and ecogenetic significance of polymorphism in the humanMGMTgene, we investigated the in-vivo characteristics of V1 and V2 methyltransferase enzyme.Esherichia colistrain KT233 (ogt−,ada−) and mer−HeLa MR cells carrying a V1 sequence exhibited almost the same level of sensitivity againstN-methyl-N′-nitro-N-nitrosoguanidine (MNNG), as did those with a wild-type sequence. The level of methyltransferase protein in those cells was essentially the same as for the wild-type and V1 samples. On the other hand,E. coliand human cells expressing V2 cDNA showed a significantly reduced level of survival. In these cells, V2 protein was hardly detected, even though mRNA was produced normally. An in-vitro translation experiment revealed that the V2 sequence had the potential to produce methyltransferase protein, as did the wild-type and V1 sequences. There was also evidence for a small amount of V2 protein being produced but rapidly degraded, thus implying that the V2 molecule is unstablein vivo. Using purified recombinant proteins, we estimated the kinetic values of wild-type and variant form of enzymes, which would support these views. From these results, we concluded that the wild-type and V1 protein have similar enzymatic and physicochemical properties, while V2 protein is considered to be unstable and rare.

 

点击下载:  PDF (378KB)



返 回