Characterization of ammonium transporters in the dinoflagellate Alexandrium catenella (Group I).

IF 2 4区生物学 Q3 MICROBIOLOGY

Fems Microbiology Letters Pub Date : 2026-01-07 DOI:10.1093/femsle/fnag120

Atsushi Kobiyama, Shinkichi Okiyama, Yuichiro Yamada

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引用次数: 0

Abstract

Alexandrium catenella (Group I) causes paralytic shellfish poisoning. Nitrogen is a critical component for both growth and toxin production in A. catenella, and recent studies have indicated that A. catenella uses ammonium as a nitrogen source. However, the molecular mechanism underlying the transportation of ammonium in A. catenella has not been identified. In this study, we attempted to isolate and characterize the ammonium transporter (AMT) in A. catenella. cDNA cloning produced a number of AMT clones. To investigate the ammonium transport capability of A. catenella AMT (AlcAMT), we selected three representative cDNAs and the clones were inserted into expression vectors. These vectors were then transferred into an amt-deficient Schizosaccharomyces pombe transgenic yeast strain, and the resulting transformants were grown on a medium containing NH4Cl as the only nitrogen source. Following transformation, only yeast transformants carrying one of the three AMTs were able to grow on the medium, confirming its ability to transport ammonium. Sequence analysis of the three AMT variants revealed that the successful variant included a K residue at sequence position 450, while the other variants contained a T residue at the same position. Mutation analysis confirmed that K450 is important for the function of AlcAMT.

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双鞭毛藻catenella Alexandrium （Group I）中铵转运体的表征。

catenella亚历山大菌（I群）引起麻痹性贝类中毒。氮是a . catenella生长和产生毒素的关键成分，最近的研究表明，a . catenella既使用无机氮，也使用铵作为氮源。然而，目前尚不清楚钩状芽孢杆菌中铵转运的分子机制。在本研究中，我们尝试分离和表征了catenella的铵转运体（AMT）。cDNA克隆产生了许多AMT克隆。为了研究catenella AMT （AlcAMT）的铵转运能力，我们选择了3个具有代表性的cdna，并将其克隆插入表达载体中。然后将这些载体转移到amt缺失的分裂糖酵母转基因酵母株中，并在含NH4Cl作为唯一氮源的培养基上生长。转化后，仅携带三种amt中的一种的酵母转化子能够在培养基上生长，证实了其运输铵的能力。对三个AMT变异的序列分析表明，成功的AMT变异在450位含有一个K残基，而其他AMT变异在相同的位置含有一个T残基。突变分析证实K450对AlcAMT的功能很重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Fems Microbiology Letters 生物-微生物学

CiteScore

4.30

自引率

0.00%

发文量

112

审稿时长

1.9 months

期刊介绍： FEMS Microbiology Letters gives priority to concise papers that merit rapid publication by virtue of their originality, general interest and contribution to new developments in microbiology. All aspects of microbiology, including virology, are covered. 2019 Impact Factor: 1.987, Journal Citation Reports (Source Clarivate, 2020) Ranking: 98/135 (Microbiology) The journal is divided into eight Sections: Physiology and Biochemistry (including genetics, molecular biology and ‘omic’ studies) Food Microbiology (from food production and biotechnology to spoilage and food borne pathogens) Biotechnology and Synthetic Biology Pathogens and Pathogenicity (including medical, veterinary, plant and insect pathogens – particularly those relating to food security – with the exception of viruses) Environmental Microbiology (including ecophysiology, ecogenomics and meta-omic studies) Virology (viruses infecting any organism, including Bacteria and Archaea) Taxonomy and Systematics (for publication of novel taxa, taxonomic reclassifications and reviews of a taxonomic nature) Professional Development (including education, training, CPD, research assessment frameworks, research and publication metrics, best-practice, careers and history of microbiology) If you are unsure which Section is most appropriate for your manuscript, for example in the case of transdisciplinary studies, we recommend that you contact the Editor-In-Chief by email prior to submission. Our scope includes any type of microorganism - all members of the Bacteria and the Archaea and microbial members of the Eukarya (yeasts, filamentous fungi, microbial algae, protozoa, oomycetes, myxomycetes, etc.) as well as all viruses.