lsm基因中编码Sm1的基序缺失会导致转录组发生明显变化,并增强嗜盐嗜碱菌细胞的群体游动活性。

Deletion of the Sm1 encoding motif in the lsm gene results in distinct changes in the transcriptome and enhanced swarming activity of Haloferax cells.

作者信息

Maier Lisa-Katharina, Benz Juliane, Fischer Susan, Alstetter Martina, Jaschinski Katharina, Hilker Rolf, Becker Anke, Allers Thorsten, Soppa Jörg, Marchfelder Anita

机构信息

Biology II, Ulm University, 89069 Ulm, Germany.

Institute of Molecular Biosciences, Goethe-University, 60438 Frankfurt, Germany.

出版信息

Biochimie. 2015 Oct;117:129-37. doi: 10.1016/j.biochi.2015.02.023. Epub 2015 Mar 6.

Abstract

Members of the Sm protein family are important for the cellular RNA metabolism in all three domains of life. The family includes archaeal and eukaryotic Lsm proteins, eukaryotic Sm proteins and archaeal and bacterial Hfq proteins. While several studies concerning the bacterial and eukaryotic family members have been published, little is known about the archaeal Lsm proteins. Although structures for several archaeal Lsm proteins have been solved already more than ten years ago, we still do not know much about their biological function, however one can confidently propose that the archaeal Lsm proteins will also be involved in RNA metabolism. Therefore, we investigated this protein in the halophilic archaeon Haloferax volcanii. The Haloferax genome encodes a single Lsm protein, the lsm gene overlaps and is co-transcribed with the gene for the ribosomal L37.eR protein. Here, we show that the reading frame of the lsm gene contains a promoter which regulates expression of the overlapping rpl37R gene. This rpl37R specific promoter ensures high expression of the rpl37R gene in exponential growth phase. To investigate the biological function of the Lsm protein we generated a lsm deletion mutant that had the coding sequence for the Sm1 motif removed but still contained the internal promoter for the downstream rpl37R gene. The transcriptome of this deletion mutant was compared to the wild type transcriptome, revealing that several genes are down-regulated and many genes are up-regulated in the deletion strain. Northern blot analyses confirmed down-regulation of two genes. In addition, the deletion strain showed a gain of function in swarming, in congruence with the up-regulation of transcripts encoding proteins required for motility.

摘要

Sm 蛋白家族成员对生命所有三个域中的细胞 RNA 代谢都很重要。该家族包括古菌和真核生物的 Lsm 蛋白、真核生物的 Sm 蛋白以及古菌和细菌的 Hfq 蛋白。虽然已经发表了几项关于细菌和真核生物家族成员的研究,但对古菌 Lsm 蛋白却知之甚少。尽管早在十多年前就已经解析了几种古菌 Lsm 蛋白的结构,但我们对它们的生物学功能仍然了解不多,不过可以有把握地推测古菌 Lsm 蛋白也会参与 RNA 代谢。因此,我们在嗜盐古菌沃氏嗜盐菌(Haloferax volcanii)中研究了这种蛋白。沃氏嗜盐菌基因组编码一种单一的 Lsm 蛋白,lsm 基因与核糖体 L37.eR 蛋白的基因重叠并共转录。在这里,我们表明 lsm 基因的阅读框包含一个启动子,该启动子调节重叠的 rpl37R 基因的表达。这个 rpl37R 特异性启动子确保 rpl37R 基因在指数生长期高表达。为了研究 Lsm 蛋白的生物学功能,我们构建了一个 lsm 缺失突变体,该突变体去除了 Sm1 基序的编码序列,但仍然包含下游 rpl37R 基因的内部启动子。将该缺失突变体的转录组与野生型转录组进行比较,发现缺失菌株中有几个基因下调,许多基因上调。Northern 印迹分析证实了两个基因的下调。此外,缺失菌株在群体游动方面表现出功能增强,这与编码运动所需蛋白质的转录本上调一致。

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