Characterization and expression of two genes encoding isoforms of a putative Na, K-ATPase in the chytridiomycete Blastocladiella emersonii
Characterization and expression of two genes encoding isoforms of a putative Na, K-ATPase in the chytridiomycete Blastocladiella emersonii
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DOI：
10.1016/s0167-4781(02)00297-x
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发表时间：
2002-06-07
期刊：
影响因子：
--
通讯作者：
Gomes, SL
中科院分区：
文献类型：
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作者：
Fietto, LG;Pugliese, L;Gomes, SL
最近从水生真菌Emersonii中发现了一个P型ATPase基因(BePAT1)，它与动物细胞中的Na，K-ATPase的α亚基有很高的相似性。生物物理学学报，1383(1998)183]。在本研究中，我们描述了第二个基因的特征，命名为BePAT2，并表明这两个基因具有不同的内含子-外显子结构，但编码的蛋白质具有超过90%的氨基酸同源性。Northern印迹和多重逆转录聚合酶链式反应(RT-PCR)分析表明，BePAT1和BePAT2基因在艾氏芽孢杆菌的生活史中具有非协调的、发育调节的表达。用免疫纯化的酶进行的磷酸酶形成实验表明，存在类似Na，K-ATPase的活性。更远-更多。利用B.Emersonii游动孢子进行的免疫荧光研究定位了这些细胞质膜上的ATPase。(C)2002 Elsevier Science B.V.保留所有权利。
A P-type ATPase gene (BePAT1) from the aquatic fungus Blastocladiella emersonii, which surprisingly showed high similarity with the a-subunit of Na, K-ATPases from animal cells, has been reported recently [Biochim. Biophys, Acta 1383 (1998) 183]. In the present study, we describe the characterization of a second gene, denominated BePAT2, and show that these two genes have a different intron-exon structure but encode putative proteins with greater than 90% amino acid identity. Northern blot and multiplex reverse transcription and polymerase chain reaction (RT-PCR) assays have revealed that BePAT1 and BePAT2 genes have a non-coordinate, developmentally regulated expression during B. emersonii life cycle. Phosphoenzyme formation experiments using the immunopurified enzymes have indicated the presence of a Na, K-ATPase-like activity. Further-more. immunofluorescence studies using B. emersonii zoospores localized the ATPases on the plasma membrane of these cells. (C) 2002 Elsevier Science B.V. All rights reserved.