摘要
目的:蛋白磷酸酶-2A(PP-2A)是真核生物中最重要的丝氨酸/苏氨酸磷酸酶之一。PP-2A的全酶由三个亚基组成:支架A亚基、催化C亚基、调节B亚基。虽然A和C亚基都由两种不同的基因编码,B亚基存在与26甚至更多的同工型,由至少15个不同的基因编码。过去的研究已经表明除了调节特异性PP-2A活性之外,各种B亚基可以具有其他功能。为了探索PP-2A的调节亚基在脊椎动物发育中的可能作用,我们克隆了编码金鱼角质素的基因,其是用PP-2A的B家族调节亚基的成员,并且确定他们的组织特异性和时间表达模式。 方法:以RT-PCR为基础的RACE进行cDNA克隆,使用RT-PCR分析金鱼角质形成细胞的mRNA表达水平。使用Western印迹分析测定来自金鱼的角蛋白表达水平。用U扫描软件进行mRNA和蛋白质表达水平的半定量。 结果:我们的研究揭示了条纹的全长cDNA由2965bp编码769个氨基酸的推导蛋白质,其与来自其他物种的同源蛋白具有非常高水平的氨基酸序列同一性。角蛋白mRNA在肾中高度表达,在脑、鳍、肌肉、肝、卵巢和鳃中较少程度,在睾丸和心脏中最低。在上述9种组织中检测到相似模式的蛋白质表达。在金鱼的发育过程中,条纹mRNA在2-细胞,多细胞和blastula阶段维持相对高的水平。然后,它基本上在原肠阶段下降并且在接下来的8个不同阶段中在该水平附近波动。在蛋白质水平,纹状体从2细胞到胃腔阶段保持较高水平,然后在神经和眼囊泡阶段减少,并逐渐增加到在眼睛色素沉着阶段达到峰值,然后在接下来的几个发展阶段略有减少。 结论:我们的研究结果表明,条纹可能在调控金鱼发展和成人组织体内平衡发挥重要作用。虽然前一功能可能通过PP-2A功能发生或可能不发生,但后一功能似乎通过PP-2A活动发生。
关键词: PP2A,调节B子成员,Striatin基因,动态平衡
Current Molecular Medicine
Title:Molecular Cloning and Developmental Expression Patterns of the Striatin Gene Encoding A Member of the Regulatory Subunits for the Protein Serine/Threonine Phosphatase-2A in Fish
Volume: 16 Issue: 10
关键词: PP2A,调节B子成员,Striatin基因,动态平衡
摘要: Purpose: The protein phosphatase-2A (PP-2A) is one of the most important serine/threonine phosphatases in eukaryotes. The holoenzyme of PP-2A consists of three subunits: a scaffold A subunit, a catalytic C subunit and a regulatory B subunit. While both A and C subunits are coded by two different genes, the B subunits exist in 26 or more isoforms which are encoded by at least 15 different genes. Previous studies have shown that besides regulating specific PP-2A activity, various B subunits may have other functions. To explore the possible roles of the regulatory subunits of PP-2A in vertebrate development, we have cloned the gene encoding goldfish striatin, a member of the B’” family regulatory subunits for PP-2A, and determined their tissue-specific and temporal expression patterns.
Methods: The cDNA cloning was conducted with RT-PCR-based RACE. The mRNA expression levels for the goldfish striatin were analyzed with RT-PCR. The expression levels of the striatin protein from goldfish were determined with Western blot analysis. The semi-quantitation of the mRNA and protein expression levels was conducted with the software of U-scanning. Results: Our study revealed that the full length cDNA for striatin consists of 2965 bp coding for a deduced protein of 769 amino acids, which bears a very high level of amino acid sequence identity with the homolog protein from other species. The striatin mRNA is highly expressed in the kidney, to a less degree in brain, fin, muscle, liver, ovary and gill, and the lowest in testis and heart. Similar pattern of protein expression is detected in the above 9 tissues. During the development of goldfish, the striatin mRNA maintains a relatively high level at the 2-cell, multiple cell and blastula stages. Then, it drops down substantially at gastrula stage and fluctuates around this level in the next 8 different stages. At the protein level, the striatin maintained higher level from 2-cell to gastrula stages, then decreased at neurula and optic vesicle stages, and gradually increased again to peak at eye pigmentation stage, then slightly decreased in the next few stages of development. Conclusions: Our results suggest that the striatin may play an important role in regulating goldfish development and adult tissue homeostasis. While the former function may or may not occur through PP- 2A functions, the later function appears to occur via PP-2A activity.Export Options
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Cite this article as:
Molecular Cloning and Developmental Expression Patterns of the Striatin Gene Encoding A Member of the Regulatory Subunits for the Protein Serine/Threonine Phosphatase-2A in Fish, Current Molecular Medicine 2016; 16 (10) . https://dx.doi.org/10.2174/1566524017666170109130230
DOI https://dx.doi.org/10.2174/1566524017666170109130230 |
Print ISSN 1566-5240 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5666 |
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