Knockout and functional analysis of two DExD/H-box family helicase genes in Sulfolobus islandicus REY15A

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• 作者：Xueguo Song ; Qihong Huang ; Jinfeng Ni ; Yang Yu ; Yulong Shen
• 刊名：Extremophiles
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：20
• 期：4
• 页码：537-546
• 全文大小：2,414 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Ecology
  
• 出版者：Springer Japan
• ISSN：1433-4909
• 卷排序：20

文摘

DExD/H-box helicases represent the largest family of helicases. They belong to superfamily 2 helicases and participate in nucleotide metabolism, ribosome biogenesis, and nucleocytoplasmic transport. The biochemical properties and structures of some DExD/H-box helicases in the archaea have been documented, but many of them have not been characterized; and reports on in vivo functional analyses are limited. In this study, we attempted gene knockout of 8 putative DExD/H-box helicases in Sulfolobus islandicus REY15A and obtained two deletion mutants, SiRe_0681 and SiRe_1605. We determined that ΔSiRe_0681 grew faster than wild type cells in the presence of methyl methanesulfonate (MMS). Flow cytometry analysis showed that this strain had fewer G1/S phase cells than the wild type, and the genes coding for cell division proteins were up-regulated. The stain ΔSiRe_1605 was more sensitive to MMS than the wild type cell, and many nucleotide metabolism and DNA repair enzymes were found to be down-regulated. Intriguingly, deletion of either gene led to silencing simultaneously of over 80 genes located at a specific region. This study provides a novel insight into the in vivo functions of predicted DExD/H-box family helicases in the archaea.KeywordsArchaeaSulfolobus islandicus REY15ADExD/H-box helicasesGene knockoutIn vivo functions