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[曲霉菌中基因表达的调控]

[Regulation of gene expression in Aspergilli].

作者信息

Tsukagoshi N

机构信息

Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-0814, Japan.

出版信息

Nihon Ishinkin Gakkai Zasshi. 1998;39(2):85-90. doi: 10.3314/jjmm.39.85.

DOI:10.3314/jjmm.39.85
PMID:9580033
Abstract

The Aspergillus oryzae Taka-amylase A (TAA) gene has been used as a model gene to characterize the regulatory mechanisms of gene expression in Aspergilli. TAA gene contained a typical eukaryotic promoter with a TATA box and putative regulatory elements such as a CCAAT sequence in its 5'-noncoding region. A nuclear protein designated AnCP bound to the CCAAT sequence. Replacement of the CCAAT sequence with CGTAA was found to abolish the binding of AnCP and to have an inhibitory effect on taa promoter activity. Although AnCP was not purified to homogeneity, AnCP appeared to have an apparent molecular mass of approximately 120 kDa. In Saccharomyces cerevisiae, the CCAAT-binding HAP complex is a heteromeric protein comprising at least four subunits (yHAP2, yHAP3, yHAP4 and yHAP5) and is involved in the regulation of genes responsible for oxidative phosphorylation. A gene designated hapC with significant homology to yHAP3 has been isolated from A. nidulans by Hynes et al. We expressed hapC gene as a fusion protein with MalE in E. coli, purified HapC protein and prepared anti-HapC antiserum. The MalE-HapC fusion protein was able to substitute for the authentic HapC in AnCP. Furthermore, addition of the anti-HapC antiserum to the DNA binding reaction mixture retarded the mobility of the shifted band. These indicate clearly that HapC protein acts as a subunit of AnCP and that AnCP is a counterpart of the yeast Hap complex.

摘要

米曲霉Taka淀粉酶A(TAA)基因已被用作模型基因来表征曲霉属中基因表达的调控机制。TAA基因在其5'非编码区包含一个带有TATA盒的典型真核启动子以及推定的调控元件,如CCAAT序列。一种名为AnCP的核蛋白与CCAAT序列结合。发现用CGTAA取代CCAAT序列会消除AnCP的结合,并对taa启动子活性产生抑制作用。尽管AnCP未被纯化至同质,但AnCP的表观分子量似乎约为120 kDa。在酿酒酵母中,CCAAT结合HAP复合物是一种异源蛋白,至少由四个亚基(yHAP2、yHAP3、yHAP4和yHAP5)组成,并参与负责氧化磷酸化的基因的调控。Hynes等人从构巢曲霉中分离出了一个与yHAP3具有显著同源性的基因hapC。我们在大肠杆菌中将hapC基因表达为与MalE的融合蛋白,纯化了HapC蛋白并制备了抗HapC抗血清。MalE-HapC融合蛋白能够替代AnCP中的真实HapC。此外,向DNA结合反应混合物中加入抗HapC抗血清会使迁移带的迁移率减慢。这些清楚地表明HapC蛋白作为AnCP的一个亚基起作用,并且AnCP是酵母Hap复合物的对应物。

相似文献

1
[Regulation of gene expression in Aspergilli].[曲霉菌中基因表达的调控]
Nihon Ishinkin Gakkai Zasshi. 1998;39(2):85-90. doi: 10.3314/jjmm.39.85.
2
The Aspergillus nidulans CCAAT-binding factor AnCP/AnCF is a heteromeric protein analogous to the HAP complex of Saccharomyces cerevisiae.构巢曲霉CCAAT结合因子AnCP/AnCF是一种异源蛋白,类似于酿酒酵母的HAP复合物。
Mol Gen Genet. 1998 Feb;257(4):404-11. doi: 10.1007/s004380050664.
3
An Aspergillus nidulans nuclear protein, AnCP, involved in enhancement of Taka-amylase A gene expression, binds to the CCAAT-containing taaG2, amdS, and gatA promoters.一种参与增强米曲霉淀粉酶A基因表达的构巢曲霉核蛋白AnCP,可与含有CCAAT的taaG2、amdS和gatA启动子结合。
Mol Gen Genet. 1997 Mar 26;254(2):119-26. doi: 10.1007/s004380050399.
4
An Aspergillus oryzae CCAAT-binding protein, AoCP, is involved in the high-level expression of the Taka-amylase A gene.米曲霉CCAAT结合蛋白AoCP参与了米曲霉淀粉酶A基因的高水平表达。
Curr Genet. 2000 Jun;37(6):380-7. doi: 10.1007/s002940000125.
5
The hapC gene of Aspergillus nidulans is involved in the expression of CCAAT-containing promoters.构巢曲霉的hapC基因参与含CCAAT启动子的表达。
Mol Gen Genet. 1996 Jun 24;251(4):412-21. doi: 10.1007/BF02172369.
6
No factors except for the hap complex increase the Taka-amylase A gene expression by binding to the CCAAT sequence in the promoter region.除了hap复合体之外,没有其他因子通过与启动子区域的CCAAT序列结合来增加 taka -淀粉酶A基因的表达。
Biosci Biotechnol Biochem. 2001 Oct;65(10):2340-2. doi: 10.1271/bbb.65.2340.
7
Aspergillus nidulans nuclear proteins bind to a CCAAT element and the adjacent upstream sequence in the promoter region of the starch-inducible Taka-amylase A gene.构巢曲霉核蛋白与淀粉诱导型淀粉酶A基因启动子区域的CCAAT元件及相邻的上游序列结合。
Mol Gen Genet. 1993 Feb;237(1-2):251-60. doi: 10.1007/BF00282807.
8
The Aspergillus nidulans multimeric CCAAT binding complex AnCF is negatively autoregulated via its hapB subunit gene.构巢曲霉多聚体CCAAT结合复合物AnCF通过其hapB亚基基因进行负向自我调节。
J Mol Biol. 2001 Mar 2;306(4):643-53. doi: 10.1006/jmbi.2001.4412.
9
Regulation of the amylolytic and (hemi-)cellulolytic genes in aspergilli.曲霉中淀粉分解和(半)纤维素分解基因的调控
J Gen Appl Microbiol. 2001 Feb;47(1):1-19. doi: 10.2323/jgam.47.1.
10
The CCAAT-binding complex of eukaryotes: evolution of a second NLS in the HapB subunit of the filamentous fungus Aspergillus nidulans despite functional conservation at the molecular level between yeast, A.nidulans and human.真核生物的CCAAT结合复合体:尽管丝状真菌构巢曲霉的HapB亚基中的第二个核定位信号(NLS)在酵母、构巢曲霉和人类之间存在分子水平的功能保守性,但它仍发生了进化。
J Mol Biol. 2005 Sep 23;352(3):517-33. doi: 10.1016/j.jmb.2005.06.068.

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