Structured mRNA induces the ribosome into a hyper-rotated state

ORAL

Abstract

During protein synthesis, mRNA and tRNA are moved through the ribosome by the process of translocation. The small diameter of the mRNA entrance tunnel only permits unstructured mRNA to pass through. However, there are structured elements within mRNA that present a barrier for translocation that must be unwound. The ribosome has been shown to unwind RNA in the absence of additional factors, but the mechanism remains unclear. Here, we show using single molecule F\"{o}rster resonance energy transfer and small angle X-ray scattering experiments a new global conformational state of the ribosome. In the presence of the frameshift inducing dnaX hairpin, we observed that the ribosomal subunits were driven into a hyper-rotated state and the L1 stalk was predominantly in an open conformation. This previously unobserved conformational state provides structural insight into the helicase activity of the ribosome and may have important implications for understanding the mechanism of reading frame maintenance.

Authors

  • Peter Cornish

    Univ of Missouri - Columbia, University of Missouri

  • Mark Neubauer

    University of Missouri-Columbia, Southern Illinois University Carbondale, Indiana University, Purdue University, Argonne National Laboratory, University of Missouri, College of Physics Science, Qingda University, Qingdao, 266071, China, Indian Institute of Science, Bangalore, India, University of Massachusetts Amherst, Iowa State University, Technical University of Denmark, University of Missouri - Columbia, University of California - San Diego, La Jolla, CA 92093, Department of Physics, University of Missouri-Columbia, Columbia, Missouri 65211, Department of Physics and Astronomy, University of Missouri-Columbia, University of Missouri, Columbia, MO, Department of Physics and Department of Biochemistry, University of Missouri, Columbia, MO 65211, Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599-3290, Univ of Missouri - Columbia, Duke University, Shanghai Jiaotong University, Ames Laboratory, U.S. DOE, Texas Center of Superconductivity and the Department of Physics, University of Houston, Institute of Physics, Siberian Division, Russian Academy of Sciences, Krasnoyarsk, 66036, Russia, Oak Ridge National Laboratory, NIST Center for Neutron Research, MU Research Reactor, Ames Laboratory and Dep. of Physics and Astronomy, Iowa State University, AmesAmes Laboratory and Dep. of Physics and Astronomy, Iowa State University, HFIR, Oak Ridge National Laboratory, University of Illinois at Urbana-Champaign

  • Mark Neubauer

    University of Missouri-Columbia, Southern Illinois University Carbondale, Indiana University, Purdue University, Argonne National Laboratory, University of Missouri, College of Physics Science, Qingda University, Qingdao, 266071, China, Indian Institute of Science, Bangalore, India, University of Massachusetts Amherst, Iowa State University, Technical University of Denmark, University of Missouri - Columbia, University of California - San Diego, La Jolla, CA 92093, Department of Physics, University of Missouri-Columbia, Columbia, Missouri 65211, Department of Physics and Astronomy, University of Missouri-Columbia, University of Missouri, Columbia, MO, Department of Physics and Department of Biochemistry, University of Missouri, Columbia, MO 65211, Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599-3290, Univ of Missouri - Columbia, Duke University, Shanghai Jiaotong University, Ames Laboratory, U.S. DOE, Texas Center of Superconductivity and the Department of Physics, University of Houston, Institute of Physics, Siberian Division, Russian Academy of Sciences, Krasnoyarsk, 66036, Russia, Oak Ridge National Laboratory, NIST Center for Neutron Research, MU Research Reactor, Ames Laboratory and Dep. of Physics and Astronomy, Iowa State University, AmesAmes Laboratory and Dep. of Physics and Astronomy, Iowa State University, HFIR, Oak Ridge National Laboratory, University of Illinois at Urbana-Champaign

  • Mark Neubauer

    University of Missouri-Columbia, Southern Illinois University Carbondale, Indiana University, Purdue University, Argonne National Laboratory, University of Missouri, College of Physics Science, Qingda University, Qingdao, 266071, China, Indian Institute of Science, Bangalore, India, University of Massachusetts Amherst, Iowa State University, Technical University of Denmark, University of Missouri - Columbia, University of California - San Diego, La Jolla, CA 92093, Department of Physics, University of Missouri-Columbia, Columbia, Missouri 65211, Department of Physics and Astronomy, University of Missouri-Columbia, University of Missouri, Columbia, MO, Department of Physics and Department of Biochemistry, University of Missouri, Columbia, MO 65211, Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599-3290, Univ of Missouri - Columbia, Duke University, Shanghai Jiaotong University, Ames Laboratory, U.S. DOE, Texas Center of Superconductivity and the Department of Physics, University of Houston, Institute of Physics, Siberian Division, Russian Academy of Sciences, Krasnoyarsk, 66036, Russia, Oak Ridge National Laboratory, NIST Center for Neutron Research, MU Research Reactor, Ames Laboratory and Dep. of Physics and Astronomy, Iowa State University, AmesAmes Laboratory and Dep. of Physics and Astronomy, Iowa State University, HFIR, Oak Ridge National Laboratory, University of Illinois at Urbana-Champaign