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FluG影响黑曲霉菌落的分泌。

FluG affects secretion in colonies of Aspergillus niger.

作者信息

Wang Fengfeng, Krijgsheld Pauline, Hulsman Marc, de Bekker Charissa, Müller Wally H, Reinders Marcel, de Vries Ronald P, Wösten Han A B

机构信息

Microbiology and Kluyver Centre for Genomics of Industrial Fermentation, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.

出版信息

Antonie Van Leeuwenhoek. 2015 Jan;107(1):225-40. doi: 10.1007/s10482-014-0321-2. Epub 2014 Nov 5.

Abstract

Colonies of Aspergillus niger are characterized by zonal heterogeneity in growth, sporulation, gene expression and secretion. For instance, the glucoamylase gene glaA is more highly expressed at the periphery of colonies when compared to the center. As a consequence, its encoded protein GlaA is mainly secreted at the outer part of the colony. Here, multiple copies of amyR were introduced in A. niger. Most transformants over-expressing this regulatory gene of amylolytic genes still displayed heterogeneous glaA expression and GlaA secretion. However, heterogeneity was abolished in transformant UU-A001.13 by expressing glaA and secreting GlaA throughout the mycelium. Sequencing the genome of UU-A001.13 revealed that transformation had been accompanied by deletion of part of the fluG gene and disrupting its 3' end by integration of a transformation vector. Inactivation of fluG in the wild-type background of A. niger also resulted in breakdown of starch under the whole colony. Asexual development of the ∆fluG strain was not affected, unlike what was previously shown in Aspergillus nidulans. Genes encoding proteins with a signal sequence for secretion, including part of the amylolytic genes, were more often downregulated in the central zone of maltose-grown ∆fluG colonies and upregulated in the intermediate part and periphery when compared to the wild-type. Together, these data indicate that FluG of A. niger is a repressor of secretion.

摘要

黑曲霉的菌落具有生长、孢子形成、基因表达和分泌方面的区域异质性。例如,与菌落中心相比,糖化酶基因glaA在菌落周边的表达水平更高。因此,其编码的蛋白GlaA主要在菌落外部分泌。在此,将amyR的多个拷贝导入黑曲霉。大多数过表达这个淀粉分解基因调控基因的转化子仍然表现出glaA表达和GlaA分泌的异质性。然而,在转化子UU-A001.13中,通过在整个菌丝体中表达glaA并分泌GlaA,异质性被消除。对UU-A001.13的基因组进行测序发现,转化过程伴随着fluG基因部分缺失以及转化载体整合破坏其3'端。在黑曲霉野生型背景中使fluG失活也导致整个菌落下方的淀粉分解。与构巢曲霉中先前所示情况不同,∆fluG菌株的无性发育不受影响。与野生型相比,编码具有分泌信号序列蛋白的基因,包括部分淀粉分解基因,在以麦芽糖为培养基生长的∆fluG菌落中心区域更常下调,而在中间部分和周边上调。这些数据共同表明,黑曲霉的FluG是分泌的抑制因子。

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