删除多酮合成酶编码基因 pks1 可阻止嗜极真菌南极隐球菌的黑色化。

IF 3.7 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
IUBMB Life Pub Date : 2024-07-16 DOI:10.1002/iub.2895
Ilaria Catanzaro, Ruben Gerrits, Ines Feldmann, Anna A. Gorbushina, Silvano Onofri, Julia Schumacher
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引用次数: 0

摘要

南极隐球菌是南极洲特有的一种黑色隐球菌,它能忍受与太空一样严酷的环境条件。特别是,它之所以能够抵御电离辐射,是因为它具有厚而高度黑色化的细胞壁,根据先前的一项调查,细胞壁可能同时含有 1,8-二羟基萘(DHN)和 L-3,4-二羟基苯丙氨酸(L-DOPA)黑色素。在南极海龟的基因组中发现了可能参与合成 DHN 黑色素的基因。其中最重要的是编码非还原型多酮合成酶(PKS)的 capks1,它是栖息于岩石中的真菌 Knufia petricola 中功能特征明显的 kppks1 的直向同源物。在酿酒酵母中,CaPKS1 或 KpPKS1 与 4'-phosphopantetheinyl 转移酶共同表达,可形成淡黄色色素,这表明 CaPKS1 是提供 DHN 黑色素前体的酶。为了研究南极藻细胞外壁黑色素层的组成和功能,研究人员通过 CRISPR/Cas9 介导的基因组编辑生成了非黑色素化突变体。尽管Δcapks1生长缓慢(长达数月),但还是获得了三个独立的非黑色素化Δcapks1突变体。突变体的生长与野生型相似,并呈现淡粉色色素沉着,这可能是类胡萝卜素所致。有趣的是,可见光对黑色素化的野生型和非黑色素化的Δcapks1菌株的生长都有不利影响。表达 H2B-GFP 融合蛋白的突变体可以通过荧光显微镜检测到光,这进一步证明了光可以穿过黑色素化的细胞壁。总之,该研究首次报道了对南极藻的遗传操作,产生了非黑色化突变体,并证明黑色素是 DHN 型的。这些突变体将有助于阐明黑色素化与在自然栖息地和太空极端条件下生存的相关性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Deletion of the polyketide synthase-encoding gene pks1 prevents melanization in the extremophilic fungus Cryomyces antarcticus

Deletion of the polyketide synthase-encoding gene pks1 prevents melanization in the extremophilic fungus Cryomyces antarcticus

Cryomyces antarcticus, a melanized cryptoendolithic fungus endemic to Antarctica, can tolerate environmental conditions as severe as those in space. Particularly, its ability to withstand ionizing radiation has been attributed to the presence of thick and highly melanized cell walls, which—according to a previous investigation—may contain both 1,8-dihydroxynaphthalene (DHN) and L-3,4 dihydroxyphenylalanine (L-DOPA) melanin. The genes putatively involved in the synthesis of DHN melanin were identified in the genome of C. antarcticus. Most important is capks1 encoding a non-reducing polyketide synthase (PKS) and being the ortholog of the functionally characterized kppks1 from the rock-inhabiting fungus Knufia petricola. The co-expression of CaPKS1 or KpPKS1 with a 4′-phosphopantetheinyl transferase in Saccharomyces cerevisiae resulted in the formation of a yellowish pigment, suggesting that CaPKS1 is the enzyme providing the precursor for DHN melanin. To dissect the composition and function of the melanin layer in the outer cell wall of C. antarcticus, non-melanized mutants were generated by CRISPR/Cas9-mediated genome editing. Notwithstanding its slow growth (up to months), three independent non-melanized Δcapks1 mutants were obtained. The mutants exhibited growth similar to the wild type and a light pinkish pigmentation, which is presumably due to carotenoids. Interestingly, visible light had an adverse effect on growth of both melanized wild-type and non-melanized Δcapks1 strains. Further evidence that light can pass the melanized cell walls derives from a mutant expressing a H2B-GFP fusion protein, which can be detected by fluorescence microscopy. In conclusion, the study reports on the first genetic manipulation of C. antarcticus, resulting in non-melanized mutants and demonstrating that the melanin is rather of the DHN type. These mutants will allow to elucidate the relevance of melanization for surviving extreme conditions found in the natural habitat as well as in space.

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来源期刊
IUBMB Life
IUBMB Life 生物-生化与分子生物学
CiteScore
10.60
自引率
0.00%
发文量
109
审稿时长
4-8 weeks
期刊介绍: IUBMB Life is the flagship journal of the International Union of Biochemistry and Molecular Biology and is devoted to the rapid publication of the most novel and significant original research articles, reviews, and hypotheses in the broadly defined fields of biochemistry, molecular biology, cell biology, and molecular medicine.
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