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首页> 外文期刊>Molecular and Cellular Biology >Gene Codon Composition Determines Differentiation-Dependent Expression of a Viral Capsid Gene in Keratinocytes In Vitro and In Vivo
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Gene Codon Composition Determines Differentiation-Dependent Expression of a Viral Capsid Gene in Keratinocytes In Vitro and In Vivo

机译：基因密码子组成决定了体外和体内角化细胞中病毒衣壳基因的差异依赖性表达。

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摘要

By establishing mouse primary keratinocytes (KCs) in culture, we were able, for the first time, to express papillomavirus major capsid (L1) proteins by transient transfection of authentic or codon-modified L1 gene expression plasmids. We demonstrate in vitro and in vivo that gene codon composition is in part responsible for differentiation-dependent expression of L1 protein in KCs. L1 mRNA was present in similar amounts in differentiated and undifferentiated KCs transfected with authentic or codon-modified L1 genes and had a similar half-life, demonstrating that L1 protein production is posttranscriptionally regulated. We demonstrate further that KCs substantially change their tRNA profiles upon differentiation. Aminoacyl-tRNAs from differentiated KCs but not undifferentiated KCs enhanced the translation of authentic L1 mRNA, suggesting that differentiation-associated change to tRNA profiles enhances L1 expression in differentiated KCs. Thus, our data reveal a novel mechanism for regulation of gene expression utilized by a virus to direct viral capsid protein expression to the site of virion assembly in mature KCs. Analysis of two structural proteins of KCs, involucrin and keratin 14, suggests that translation of their mRNAs is also regulated, in association with KC differentiation in vitro, by a similar mechanism.
机译：通过在培养物中建立小鼠原代角质形成细胞（KCs），我们能够通过瞬时转染真实或经密码子修饰的L1基因表达质粒来首次表达乳头瘤病毒主要衣壳（L1）蛋白。我们在体外和体内证明基因密码子组成部分负责KCs中L1蛋白的分化依赖性表达。 L1 mRNA在以真实或经密码子修饰的L1基因转染的分化和未分化KC中以相似的量存在，并且具有相似的半衰期，表明L1蛋白的生产受转录后调控。我们进一步证明，KCs在分化后会实质上改变其tRNA谱。来自分化的KC而不是未分化的KC的氨酰基tRNA增强了真实L1 mRNA的翻译，这表明与分化相关的tRNA图谱的改变增强了分化KC中L1的表达。因此，我们的数据揭示了一种调节基因表达的新机制，该机制利用病毒将病毒衣壳蛋白表达引导至成熟KC中的病毒体装配位点。对KCs的两个结构蛋白（involucrin和keratin 14）的分析表明，与KC体外分化相关，它们的mRNA的翻译也受到类似机制的调节。

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