Cao Cunwei, Liu Wei, Li Ruoyu
Department of Dermatology, Research Center for Medical Mycology, Peking University First Hospital, Peking University, No. 8, Xishiku St., West District, 100034 Beijing, People's Republic of China.
Mycopathologia. 2009 Jul;168(1):23-30. doi: 10.1007/s11046-009-9192-x. Epub 2009 Mar 18.
Penicillium marneffei is an intracellular fungal pathogen. Being resistant to the reactive oxygen species (ROS) produced by phagocytes is a key step to the surviving of P. marneffei within the phagocytes, as well as its invasion to the host. SKN7, a transcription factor, contributes to the oxidative stress response in yeasts. We cloned the SKN7 ortholog in P. marneffei and compared its transcription on both mycelial and yeast phase, identified its function by complement the S. cerevisiae skn7 disruptant strain. The result showed the full sequence of SKN7 gene was about 2.5 kb, open reading frame extended to 1,845 bp and encoded a putative protein of 614 amino acids. Comparative analysis of the nucleotide sequence of genomic DNA and cDNA confirmed the presence of four introns and two highly conserved HSF-DNA-bind and REC domain. The deduced amino acid sequence was homologous to SKN7 from other fungi. Further, P. marneffei cDNA can partly complement S. cerevisiae skn7 disruptant strain, which was not viable in the presence of 2.5 mM H2O2. The expression level of SKN7 on the two phases, however, had no difference. These results indicated that P. marneffei SKN7 could response to oxidative stress and it was not a phase-specific gene.
马尔尼菲青霉是一种细胞内真菌病原体。对吞噬细胞产生的活性氧(ROS)具有抗性是马尔尼菲青霉在吞噬细胞内存活及其侵入宿主的关键步骤。SKN7是一种转录因子,有助于酵母中的氧化应激反应。我们克隆了马尔尼菲青霉中的SKN7直系同源基因,比较了其在菌丝体和酵母阶段的转录情况,并通过补充酿酒酵母skn7缺失菌株来鉴定其功能。结果表明,SKN7基因的全长序列约为2.5 kb,开放阅读框延伸至1845 bp,编码一个推定的614个氨基酸的蛋白质。对基因组DNA和cDNA核苷酸序列的比较分析证实存在四个内含子以及两个高度保守的热休克因子-DNA结合域和REC结构域。推导的氨基酸序列与其他真菌的SKN7同源。此外,马尔尼菲青霉cDNA可以部分补充酿酒酵母skn7缺失菌株,该菌株在2.5 mM H2O2存在下无法存活。然而,SKN7在两个阶段的表达水平没有差异。这些结果表明,马尔尼菲青霉SKN7可以对氧化应激作出反应,并且它不是一个阶段特异性基因。