Pregnenolone esterification in Saccharomyces cerevisiae -: A potential detoxification mechanism

被引:44
作者
Cauet, G
Degryse, E
Ledoux, C
Spagnoli, R
Achstetter, T
机构
[1] Transgene SA, F-67082 Strasbourg, France
[2] Hoechst Marion Roussel, Romainville, France
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1999年 / 261卷 / 01期
关键词
ABC transporters; alcohol acetyltransferase; detoxification; esterification; steroids;
D O I
10.1046/j.1432-1327.1999.00282.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
While studying the effect of steroids on the growth of the yeast Saccharomyces cerevisiae, we found that pregnenolone was converted into the acetate ester. This reaction was identified as a transfer of the acetyl group from acetyl-CoA to the 3 beta-hydroxyl group of pregnenolone. The corresponding enzyme, acetyl-CoA: pregnenolone acetyltransferase (APAT) is specific for Delta(5)- or Delta(4)-3 beta-hydroxysteroids and short-chain acyl-CoAs. The apparent K-m for pregnenolone is approximate to 0.5 mu M. The protein associated with APAT activity was partially purified and finally isolated from an SDS/polyacrylamide gel. Tryptic peptides were generated and N-terminally sequenced. Two peptide sequences allowed the identification of an open reading frame (YGR177c, in the S. cerevisiae genome database) translating into a 62-kDa protein of hitherto unknown function. This protein encoded by a gene known as ATF2 displays 37% identity with an alcohol acetyltransferase encoded by the yeast gene ATF1. Disruption of ATF2 led to the complete elimination of APAT activity and consequently abolished the esterification of pregnenolone. In addition, a toxic effect of pregnenolone linked to the disruption of ATF2 was observed. Pregnenolone toxicity is more pronounced when the atf 2-Delta mutation is introduced in a yeast strain devoid of the ATP-binding cassette transporters, PDR5 and SNQ2. Our results suggest that Atf2p (APAT) plays an active role in the detoxification of 3 beta-hydroxysteroids in association with the efflux pumps Pdr5p and Snq2p.
引用
收藏
页码:317 / 324
页数:8
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