Tosco A, Gargano S, Kobayashi G S, Maresca B
International Institute of Genetics and Biophysics, CNR, Naples, Italy.
Biochem Biophys Res Commun. 1997 Jan 13;230(2):457-61. doi: 10.1006/bbrc.1996.5984.
We have characterized a region of the promoter of a cloned delta9-desaturase gene (Ole1) of Histoplasma capsulatum, a dimorphic pathogenic fungus of humans. The product of the delta9-desaturase gene is involved in regulating membrane fluid state in animal cells and microorganisms. To identify sequences critical for Ole1 expression in both the saprobic mycelial and parasitic yeast phases of this organism, we performed a deletion analysis. Evidence is presented that a 240 nt region of the proximal promoter is involved in a phase-specific binding in vitro. By sequence analysis we have identified one likely regulatory element that coincides with an AP1 binding site (TGACTAA) that is located at -740 nt of 5'-upstream from the ATG. Using gel mobility shift assays, we show that this cis-acting element binds nuclear proteins extracted from the yeast and mycelial phases of H. capsulatum that may participate in control of expression of the delta9-desaturase gene.
我们已经对荚膜组织胞浆菌(一种人类双相致病真菌)克隆的Δ9-去饱和酶基因(Ole1)启动子的一个区域进行了表征。Δ9-去饱和酶基因的产物参与调节动物细胞和微生物中的膜流体状态。为了鉴定该生物体在腐生菌丝体和寄生酵母阶段中对Ole1表达至关重要的序列,我们进行了缺失分析。有证据表明,近端启动子的一个240 nt区域参与了体外阶段特异性结合。通过序列分析,我们确定了一个可能的调控元件,它与位于ATG上游5'-740 nt处的一个AP1结合位点(TGACTAA)一致。使用凝胶迁移率变动分析,我们表明这个顺式作用元件结合从荚膜组织胞浆菌的酵母和菌丝体阶段提取的核蛋白,这些核蛋白可能参与Δ9-去饱和酶基因表达的控制。