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秀丽隐杆线虫的 sirtuin 基因 sir-2.1 广泛表达,并在热量限制时被诱导。

The Caenorhabditis elegans sirtuin gene, sir-2.1, is widely expressed and induced upon caloric restriction.

机构信息

Institute of Integrative and Comparative Biology, Faculty of Biological Sciences, The University of Leeds, Woodhouse Lane, Leeds LS2 9JT, UK.

出版信息

Mech Ageing Dev. 2009 Nov-Dec;130(11-12):762-70. doi: 10.1016/j.mad.2009.10.001.

Abstract

INTRODUCTION

As in yeast, flies and mammals, over-expression of the Caenorhabditis elegans sirtuin gene sir-2.1 leads to extension of lifespan and deletion of the gene shortens lifespan. The sir-2.1 gene, however, is located in an operon, an organization not taken into account in previous studies of this gene's expression.

MATERIALS AND METHODS

Recombineering allowed insertion of both a mCherry and a gfp reporter gene precisely at the end of the two protein-coding regions of the 4.5kb sir-2.1 operon within a 29.3kb genomic DNA fosmid clone.

RESULTS AND DISCUSSION

In C. elegans transgenic for this recombineered fosmid, with abundant food, the sir-2.1::mCherry distribution indicated that sir-2.1 is indeed expressed in the hypodermis and many nerve cells, as previously described, but also in the intestine and in muscles. This broader expression of sir-2.1, which would fit with an expectation that SIR2.1 function in influencing lifespan might be required in most cell types, arises from transcription starting with the gene upstream of sir-2.1 in the operon. Importantly, the expression of both genes in the operon increases upon starvation, this induction also depending on the operon promoter. Furthermore, SIR-2.1::mCherry undergoes a dynamic subcellular relocalization through starvation.

摘要

简介

与酵母、苍蝇和哺乳动物一样,秀丽隐杆线虫中的 sir-2.1 长寿基因的过表达会延长寿命,而该基因的缺失则会缩短寿命。然而,sir-2.1 基因位于操纵子中,这在以前对该基因表达的研究中并未考虑到。

材料与方法

通过重组酶技术,我们可以将 mCherry 和 gfp 报告基因精确地插入到 4.5kb 的 sir-2.1 操纵子的两个蛋白质编码区的末端,该操纵子位于 29.3kb 的基因组 DNA fosmid 克隆中。

结果与讨论

在这种重组 fosmid 的转基因秀丽隐杆线虫中,在丰富的食物条件下,sir-2.1::mCherry 的分布表明 sir-2.1 确实在表皮和许多神经细胞中表达,这与之前的描述一致,但也在肠道和肌肉中表达。这种 sir-2.1 的更广泛表达,与 SIR2.1 影响寿命的功能可能需要在大多数细胞类型中发挥作用的预期相符,这是由于操纵子上游基因的转录起始引起的。重要的是,操纵子中两个基因的表达在饥饿时增加,这种诱导也依赖于操纵子启动子。此外,SIR-2.1::mCherry 会经历一个通过饥饿引起的动态亚细胞重定位。

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