Insights into the Structure of Dimeric RNA Helicase CsdA and Indispensable Role of Its C-Terminal Regions

Structure. 2017 Dec 5;25(12):1795-1808.e5. doi: 10.1016/j.str.2017.09.013. Epub 2017 Oct 26.

Abstract

CsdA has been proposed to be essential for the biogenesis of ribosome and gene regulation after cold shock. However, the structure of CsdA and the function of its long C-terminal regions are still unclear. Here, we solved all of the domain structures of CsdA and found two previously uncharacterized auxiliary domains: a dimerization domain (DD) and an RNA-binding domain (RBD). Small-angle X-ray scattering experiments helped to track the conformational flexibilities of the helicase core domains and C-terminal regions. Biochemical assays revealed that DD is indispensable for stabilizing the CsdA dimeric structure. We also demonstrate for the first time that CsdA functions as a stable dimer at low temperature. The C-terminal regions are critical for RNA binding and efficient enzymatic activities. CsdA_RBD could specifically bind to the regions with a preference for single-stranded G-rich RNA, which may help to bring the helicase core to unwind the adjacent duplex.

Keywords: ATPase activity; CsdA; DEAD-box helicase; RNA binding; crystal structure; dimerization domain; small-angle X-ray scattering; unwinding activity.

MeSH terms

  • Binding Sites
  • DEAD-box RNA Helicases / chemistry*
  • DEAD-box RNA Helicases / metabolism
  • Escherichia coli / enzymology
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / metabolism
  • Protein Binding
  • Protein Multimerization
  • RNA / metabolism

Substances

  • Escherichia coli Proteins
  • RNA
  • deaD protein, E coli
  • DEAD-box RNA Helicases