A hap1 mutation in a laboratory strain of Saccharomyces cerevisiae results in decreased expression of ergosterol-related genes and cellular ergosterol content compared to sake yeast

被引:38
作者
Tamura, KI
Gu, YQ
Wang, Q
Yamada, T
Ito, K
Shimoi, H
机构
[1] Hiroshima Univ, Grad Sch Adv Sci Matter, Dept Mol Biotechnol, Hiroshima 7398530, Japan
[2] Takara Shuzo Co Ltd, Otsu, Shiga 5202193, Japan
[3] Shihezi Univ, Coll Med, Mol Med Lab, Xinjiang 832002, Peoples R China
[4] Kiku Masamune Sake Brewing Co Ltd, Gen Res Lab, Higashinada Ku, Kobe, Hyogo 6580026, Japan
[5] Natl Res Inst Brewing, Hiroshima 7390046, Japan
关键词
sake yeast; HAP1; HMG1; ergosterol;
D O I
10.1016/S1389-1723(04)00260-9
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
DNA microarray and Northern blot analysis revealed that a sake yeast strain Kyokai no. 7 (K7) showed higher expression of genes encoding proteins involved in ergosterol biosynthesis than a laboratory yeast strain X2180. We hypothesized that these differences in expression levels were caused by a defect of a transcriptional factor Hap1, because strain X2180 contained a Ty1 insertion mutation in the HAP1 gene. To confirm this, we constructed a strain X2180 derivative (strain HX) that contained the wild-type HAP1 genes originating from strain K7. The expression levels of ergosterol-related genes and cellular ergosterol content in strain HX were higher than those in strain X2180 and were almost comparable to those in strain K7. These results suggest that the differences in the expression levels of ergosterol-related genes and ergosterol content between strains K7 and X2180 were largely caused by the hap1 mutation in strain X2180. Involvement of the mutated Hap1 in the differential gene expression between strain K7 and strain X2180 was further confirmed by a lacZ reporter assay of HMG1, one of the Hap1-regulated genes. We also revealed that the HMG1 promoter region between -500 and -376 was important in the transcriptional activation by Hap1.
引用
收藏
页码:159 / 166
页数:8
相关论文
共 30 条
[1]  
[Anonymous], METHOD ENZYMOL
[2]  
[Anonymous], METHOD ENZYMOL
[3]   SACCHAROMYCES-CEREVISIAE CONTAINS 2 FUNCTIONAL GENES ENCODING 3-HYDROXY-3-METHYLGLUTARYL-COENZYME-A REDUCTASE [J].
BASSON, ME ;
THORSNESS, M ;
RINE, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (15) :5563-5567
[4]   The yeast transcriptome in aerobic and hypoxic conditions:: effects of hap1, rox1, rox3 and srb10 deletions [J].
Becerra, M ;
Lombardía-Ferreira, LJ ;
Hauser, NC ;
Hoheisel, JD ;
Tizon, B ;
Cerdán, ME .
MOLECULAR MICROBIOLOGY, 2002, 43 (03) :545-555
[5]   Genetic dissection of transcriptional regulation in budding yeast [J].
Brem, RB ;
Yvert, G ;
Clinton, R ;
Kruglyak, L .
SCIENCE, 2002, 296 (5568) :752-755
[6]  
BURKE D, 2000, METHODS YEAST GENETI, P123
[7]   LARGE-SCALE ANALYSIS OF GENE-EXPRESSION, PROTEIN LOCALIZATION, AND GENE DISRUPTION SACCHAROMYCES-CEREVISIAE [J].
BURNS, N ;
GRIMWADE, B ;
ROSSMACDONALD, PB ;
CHOI, EY ;
FINBERG, K ;
ROEDER, GS ;
SNYDER, M .
GENES & DEVELOPMENT, 1994, 8 (09) :1087-1105
[8]  
Daum G, 1998, YEAST, V14, P1471, DOI 10.1002/(SICI)1097-0061(199812)14:16<1471::AID-YEA353>3.0.CO
[9]  
2-Y
[10]   FEEDBACK-REGULATION OF 3-HYDROXY-3-METHYLGLUTARYL COENZYME-A REDUCTASE IN SACCHAROMYCES-CEREVISIAE [J].
DIMSTERDENK, D ;
THORSNESS, MK ;
RINE, J .
MOLECULAR BIOLOGY OF THE CELL, 1994, 5 (06) :655-665