How RNA unfolds and refolds.

Understanding how RNA folds and what causes it to unfold has become more important as knowledge of the diverse functions of RNA has increased. Here we review the contributions of single-molecule experiments to providing answers to questions such as: How much energy is required to unfold a secondary or tertiary structure? How fast is the process? How do helicases unwind double helices? Are the unwinding activities of RNA-dependent RNA polymerases and of ribosomes different from other helicases? We discuss the use of optical tweezers to monitor the unfolding activities of helicases, polymerases, and ribosomes, and to apply force to unfold RNAs directly. We also review the applications of fluorescence and fluorescence resonance energy transfer to measure RNA dynamics.

[1]  Colin Echeverría Aitken,et al.  Translation at the single-molecule level. , 2008, Annual review of biochemistry.

[2]  C. Joo,et al.  Advances in single-molecule fluorescence methods for molecular biology. , 2008, Annual review of biochemistry.

[3]  Ignacio Tinoco,et al.  Following translation by single ribosomes one codon at a time , 2008, Nature.

[4]  Ignacio Tinoco,et al.  Characterization of the Mechanical Unfolding of RNA Pseudoknots , 2007, Journal of Molecular Biology.

[5]  I. Tinoco,et al.  NS3 helicase actively separates RNA strands and senses sequence barriers ahead of the opening fork , 2007, Proceedings of the National Academy of Sciences.

[6]  P. Barbara,et al.  Probing nucleation, reverse annealing, and chaperone function along the reaction path of HIV-1 single-strand transfer , 2007, Proceedings of the National Academy of Sciences.

[7]  X. Zhuang,et al.  Dissecting the multistep reaction pathway of an RNA enzyme by single-molecule kinetic “fingerprinting” , 2007, Proceedings of the National Academy of Sciences.

[8]  Taekjip Ha,et al.  Spring-Loaded Mechanism of DNA Unwinding by Hepatitis C Virus NS3 Helicase , 2007, Science.

[9]  Michelle D. Wang,et al.  Single-Molecule Studies Reveal Dynamics of DNA Unwinding by the Ring-Shaped T7 Helicase , 2007, Cell.

[10]  D. Wigley,et al.  Structure and mechanism of helicases and nucleic acid translocases. , 2007, Annual review of biochemistry.

[11]  K. Schulten,et al.  Single-Molecule Experiments in Vitro and in Silico , 2007, Science.

[12]  F. Ritort,et al.  Force unfolding kinetics of RNA using optical tweezers. II. Modeling experiments. , 2007, Biophysical journal.

[13]  Ignacio Tinoco,et al.  Force unfolding kinetics of RNA using optical tweezers. I. Effects of experimental variables on measured results. , 2007, Biophysical journal.

[14]  Ioulia Rouzina,et al.  Quantifying force-dependent and zero-force DNA intercalation by single-molecule stretching , 2007, Nature Methods.

[15]  Ignacio Tinoco,et al.  Real-time control of the energy landscape by force directs the folding of RNA molecules , 2007, Proceedings of the National Academy of Sciences.

[16]  A. Buhot,et al.  Stretching of homopolymeric RNA reveals single-stranded helices and base-stacking. , 2007, Physical review letters.

[17]  Carlos Bustamante,et al.  Backtracking determines the force sensitivity of RNAP II in a factor-dependent manner , 2007, Nature.

[18]  S. Nader S. Reihani,et al.  Correlation between mechanical strength of messenger RNA pseudoknots and ribosomal frameshifting , 2007, Proceedings of the National Academy of Sciences.

[19]  Wojciech Kasprzak,et al.  Bridging the gap in RNA structure prediction. , 2007, Current opinion in structural biology.

[20]  John E. Johnson,et al.  DNA packaging and delivery machines in tailed bacteriophages. , 2007, Current opinion in structural biology.

[21]  Shi-Jie Chen,et al.  Biphasic folding kinetics of RNA pseudoknots and telomerase RNA activity. , 2007, Journal of molecular biology.

[22]  Michael D. Stone,et al.  Stepwise protein-mediated RNA folding directs assembly of telomerase ribonucleoprotein , 2007, Nature.

[23]  A. Pyle,et al.  Folding of group II introns: a model system for large, multidomain RNAs? , 2007, Trends in biochemical sciences.

[24]  P. Barbara,et al.  Insights on the role of nucleic acid/protein interactions in chaperoned nucleic acid rearrangements of HIV-1 reverse transcription , 2007, Proceedings of the National Academy of Sciences.

[25]  G. Nienhaus,et al.  Mg2+-dependent folding of a Diels-Alderase ribozyme probed by single-molecule FRET analysis , 2007, Nucleic acids research.

[26]  David E Draper,et al.  Tertiary structure of an RNA pseudoknot is stabilized by "diffuse" Mg2+ ions. , 2007, Biochemistry.

[27]  H. Al‐Hashimi,et al.  Resolving fast and slow motions in the internal loop containing stem-loop 1 of HIV-1 that are modulated by Mg2+ binding: role in the kissing–duplex structural transition , 2007, Nucleic acids research.

[28]  Micah J. McCauley,et al.  Mechanisms of DNA binding determined in optical tweezers experiments. , 2007, Biopolymers.

[29]  Hui Zhang,et al.  Counting of six pRNAs of phi29 DNA‐packaging motor with customized single‐molecule dual‐view system , 2007, The EMBO journal.

[30]  Hesham H. Ali,et al.  High sensitivity RNA pseudoknot prediction , 2006, Nucleic acids research.

[31]  D. Thirumalai,et al.  Mechanical unfolding of RNA: from hairpins to structures with internal multiloops. , 2006, Biophysical journal.

[32]  S. Strobel,et al.  Structural investigation of the GlmS ribozyme bound to Its catalytic cofactor. , 2007, Chemistry & biology.

[33]  A. Pyle,et al.  Alternative roles for metal ions in enzyme catalysis and the implications for ribozyme chemistry. , 2007, Chemical reviews.

[34]  Erik Winfree,et al.  Thermodynamic Analysis of Interacting Nucleic Acid Strands , 2007, SIAM Rev..

[35]  David D. L. Minh Free-energy reconstruction from experiments performed under different biasing programs. , 2006, Physical review. E, Statistical, nonlinear, and soft matter physics.

[36]  R. Landick The regulatory roles and mechanism of transcriptional pausing. , 2006, Biochemical Society transactions.

[37]  D. Herschlag,et al.  Direct Measurement of the Full, Sequence-Dependent Folding Landscape of a Nucleic Acid , 2006, Science.

[38]  Nicolas Foloppe,et al.  Towards the discovery of drug-like RNA ligands? , 2006, Drug discovery today.

[39]  Ignacio Tinoco,et al.  Unusual mechanical stability of a minimal RNA kissing complex , 2006, Proceedings of the National Academy of Sciences.

[40]  Ignacio Tinoco,et al.  Determination of thermodynamics and kinetics of RNA reactions by force , 2006, Quarterly Reviews of Biophysics.

[41]  M. Rodnina,et al.  Role and timing of GTP binding and hydrolysis during EF-G-dependent tRNA translocation on the ribosome , 2006, Proceedings of the National Academy of Sciences.

[42]  K. Réblová,et al.  Trapped water molecules are essential to structural dynamics and function of a ribozyme , 2006, Proceedings of the National Academy of Sciences.

[43]  M. Rudolph,et al.  Crystal structure and nucleotide binding of the Thermus thermophilus RNA helicase Hera N-terminal domain. , 2006, Journal of molecular biology.

[44]  T. Ha,et al.  Unraveling helicase mechanisms one molecule at a time , 2006, Nucleic acids research.

[45]  D. Turner,et al.  A set of nearest neighbor parameters for predicting the enthalpy change of RNA secondary structure formation , 2006, Nucleic acids research.

[46]  T. Steitz Visualizing polynucleotide polymerase machines at work , 2006, The EMBO journal.

[47]  C. A. Theimer,et al.  Structure and function of telomerase RNA. , 2006, Current opinion in structural biology.

[48]  F. Ritort,et al.  Force-dependent fragility in RNA hairpins. , 2006, Physical review letters.

[49]  Michelle D. Wang,et al.  Single-molecule analysis of RNA polymerase transcription. , 2006, Annual review of biophysics and biomolecular structure.

[50]  D. Herschlag,et al.  Nanomechanical measurements of the sequence-dependent folding landscapes of single nucleic acid hairpins. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Julie L. Fiore,et al.  Metal ion dependence, thermodynamics, and kinetics for intramolecular docking of a GAAA tetraloop and receptor connected by a flexible linker. , 2006, Biochemistry.

[52]  D. Côte,et al.  Probing DNA and RNA single molecules with a double optical tweezer , 2006, The European physical journal. E, Soft matter.

[53]  Qi Zhang,et al.  Resolving the Motional Modes That Code for RNA Adaptation , 2006, Science.

[54]  Changbong Hyeon,et al.  Forced-unfolding and force-quench refolding of RNA hairpins. , 2006, Biophysical journal.

[55]  N. Piganeau,et al.  Stabilities of HIV-1 DIS type RNA loop–loop interactions in vitro and in vivo , 2006, Nucleic acids research.

[56]  I. Tinoco,et al.  RNA translocation and unwinding mechanism of HCV NS3 helicase and its coordination by ATP , 2006, Nature.

[57]  Ignacio Tinoco,et al.  Probing the mechanical folding kinetics of TAR RNA by hopping, force-jump, and force-ramp methods. , 2006, Biophysical journal.

[58]  Michelle D. Wang,et al.  Detection of high-affinity and sliding clamp modes for MSH2-MSH6 by single-molecule unzipping force analysis. , 2005, Molecular cell.

[59]  A. Jäschke,et al.  Nucleic acid enzymes. , 2005, Current opinion in biotechnology.

[60]  C. Jarzynski,et al.  Verification of the Crooks fluctuation theorem and recovery of RNA folding free energies , 2005, Nature.

[61]  Julie L. Fiore,et al.  Docking kinetics and equilibrium of a GAAA tetraloop-receptor motif probed by single-molecule FRET. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[62]  D. Lilley,et al.  Structure, folding and mechanisms of ribozymes. , 2005, Current opinion in structural biology.

[63]  D. Draper,et al.  Ions and RNA folding. , 2005, Annual review of biophysics and biomolecular structure.

[64]  X. Zhuang Single-molecule RNA science. , 2005, Annual review of biophysics and biomolecular structure.

[65]  Smita S. Patel,et al.  A Brownian motor mechanism of translocation and strand separation by hepatitis C virus helicase , 2005, Nature Structural &Molecular Biology.

[66]  S. Woodson Metal ions and RNA folding: a highly charged topic with a dynamic future. , 2005, Current opinion in chemical biology.

[67]  D. Thirumalai,et al.  Mechanical unfolding of RNA hairpins. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[68]  Xiaowei Zhuang,et al.  Single-molecule RNA folding. , 2005, Accounts of chemical research.

[69]  Gerhard Hummer,et al.  Free energy surfaces from single-molecule force spectroscopy. , 2005, Accounts of chemical research.

[70]  F. Marchesoni,et al.  Brownian motors , 2004, cond-mat/0410033.

[71]  F. Ritort,et al.  Thermodynamic and kinetic aspects of RNA pulling experiments. , 2004, Biophysical journal.

[72]  K. Musier-Forsyth,et al.  Nucleic acid chaperone activity of HIV-1 nucleocapsid protein: critical role in reverse transcription and molecular mechanism. , 2005, Progress in nucleic acid research and molecular biology.

[73]  I. Tinoco,et al.  The effect of force on thermodynamics and kinetics: unfolding single RNA molecules. , 2004, Biochemical Society transactions.

[74]  M. Rodnina,et al.  Mechanisms of elongation on the ribosome: dynamics of a macromolecular machine. , 2004, Biochemical Society transactions.

[75]  D. Lilley,et al.  Observation of internal cleavage and ligation reactions of a ribozyme , 2004, Nature Structural &Molecular Biology.

[76]  J. Puglisi,et al.  tRNA selection and kinetic proofreading in translation , 2004, Nature Structural &Molecular Biology.

[77]  Yeonee Seol,et al.  Elastic properties of a single-stranded charged homopolymeric ribonucleotide. , 2004, Physical review letters.

[78]  Steven Chu,et al.  tRNA dynamics on the ribosome during translation. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[79]  V. Serebrov,et al.  Periodic cycles of RNA unwinding and pausing by hepatitis C virus NS3 helicase , 2004, Nature.

[80]  M. Davies,et al.  Single-molecule investigations of RNA dissociation. , 2004, Biophysical journal.

[81]  A. Pyle,et al.  Backbone tracking by the SF2 helicase NPH-II , 2004, Nature Structural &Molecular Biology.

[82]  Ignacio Tinoco,et al.  Force as a useful variable in reactions: unfolding RNA. , 2004, Annual review of biophysics and biomolecular structure.

[83]  P. Linder,et al.  DEAD-box proteins: the driving forces behind RNA metabolism , 2004, Nature Reviews Molecular Cell Biology.

[84]  Christoph Flamm,et al.  Determination of thermodynamic parameters for HIV DIS type loop-loop kissing complexes. , 2004, Nucleic acids research.

[85]  T. Lohman,et al.  General methods for analysis of sequential "n-step" kinetic mechanisms: application to single turnover kinetics of helicase-catalyzed DNA unwinding. , 2003, Biophysical journal.

[86]  N. Walter,et al.  Diffusely bound Mg2+ ions slightly reorient stems I and II of the hammerhead ribozyme to increase the probability of formation of the catalytic core. , 2003, Biochemistry.

[87]  Michelle D. Wang,et al.  Dynamic force spectroscopy of protein-DNA interactions by unzipping DNA. , 2003, Physical review letters.

[88]  Michael Zuker,et al.  Mfold web server for nucleic acid folding and hybridization prediction , 2003, Nucleic Acids Res..

[89]  Ivo L. Hofacker,et al.  Vienna RNA secondary structure server , 2003, Nucleic Acids Res..

[90]  V. DeRose,et al.  Metal ion binding to catalytic RNA molecules. , 2003, Current opinion in structural biology.

[91]  Wolfgang Wintermeyer,et al.  An elongation factor G-induced ribosome rearrangement precedes tRNA-mRNA translocation. , 2003, Molecular cell.

[92]  X. Zhuang,et al.  Exploration of the transition state for tertiary structure formation between an RNA helix and a large structured RNA. , 2003, Journal of molecular biology.

[93]  Ignacio Tinoco,et al.  Identifying Kinetic Barriers to Mechanical Unfolding of the T. thermophila Ribozyme , 2003, Science.

[94]  P. V. von Hippel,et al.  Helicase mechanisms and the coupling of helicases within macromolecular machines Part II: Integration of helicases into cellular processes , 2003, Quarterly Reviews of Biophysics.

[95]  Carlos Bustamante,et al.  Optical-trap force transducer that operates by direct measurement of light momentum. , 2003, Methods in enzymology.

[96]  P. V. von Hippel,et al.  Helicase mechanisms and the coupling of helicases within macromolecular machines Part I: Structures and properties of isolated helicases , 2002, Quarterly Reviews of Biophysics.

[97]  C. Ehresmann,et al.  RNA loop-loop interactions as dynamic functional motifs. , 2002, Biochimie.

[98]  Benjamin C. Jantzen,et al.  Probing protein-DNA interactions by unzipping a single DNA double helix. , 2002, Biophysical journal.

[99]  I. Tinoco,et al.  Equilibrium Information from Nonequilibrium Measurements in an Experimental Test of Jarzynski's Equality , 2002, Science.

[100]  Paul Ahlquist,et al.  RNA-Dependent RNA Polymerases, Viruses, and RNA Silencing , 2002, Science.

[101]  I. Shih,et al.  Catalytic strategies of the hepatitis delta virus ribozymes. , 2002, Annual review of biochemistry.

[102]  D. Draper,et al.  A thermodynamic framework for Mg2+ binding to RNA , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[103]  J. Liphardt,et al.  Reversible Unfolding of Single RNA Molecules by Mechanical Force , 2001, Science.

[104]  E. Westhof,et al.  TectoRNA: modular assembly units for the construction of RNA nano-objects. , 2001, Nucleic acids research.

[105]  C. Wong,et al.  RNA as a target for small molecules. , 2000, Current opinion in chemical biology.

[106]  X. Zhuang,et al.  A single-molecule study of RNA catalysis and folding. , 2000, Science.

[107]  E. Siggia,et al.  Modeling RNA folding paths with pseudoknots: application to hepatitis delta virus ribozyme. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[108]  J. Doudna,et al.  Ribozyme structures and mechanisms. , 2000, Annual review of biochemistry.

[109]  D. Giedroc,et al.  Structure, stability and function of RNA pseudoknots involved in stimulating ribosomal frameshifting1 , 2000, Journal of Molecular Biology.

[110]  A. Pyle,et al.  The DExH protein NPH-II is a processive and directional motor for unwinding RNA , 2000, Nature.

[111]  I. Tinoco,et al.  How RNA folds. , 1999, Journal of molecular biology.

[112]  X. Zhuang,et al.  Ligand-induced conformational changes observed in single RNA molecules. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[113]  G. Crooks Entropy production fluctuation theorem and the nonequilibrium work relation for free energy differences. , 1999, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[114]  E Rivas,et al.  A dynamic programming algorithm for RNA structure prediction including pseudoknots. , 1998, Journal of molecular biology.

[115]  T. Pape,et al.  Complete kinetic mechanism of elongation factor Tu‐dependent binding of aminoacyl‐tRNA to the A site of the E.coli ribosome , 1998, The EMBO journal.

[116]  J P Griffith,et al.  Hepatitis C virus NS3 RNA helicase domain with a bound oligonucleotide: the crystal structure provides insights into the mode of unwinding. , 1998, Structure.

[117]  Anders Liljas,et al.  Structural aspects of protein synthesis , 2004, Nature Structural Biology.

[118]  E. Evans,et al.  Dynamic strength of molecular adhesion bonds. , 1997, Biophysical journal.

[119]  C. Jarzynski Nonequilibrium Equality for Free Energy Differences , 1996, cond-mat/9610209.

[120]  C. Kundrot,et al.  Crystal Structure of a Group I Ribozyme Domain: Principles of RNA Packing , 1996, Science.

[121]  B Honig,et al.  Salt effects on polyelectrolyte–ligand binding: Comparison of Poisson–Boltzmann, and limiting law/counterion binding models , 1995, Biopolymers.

[122]  B Honig,et al.  Salt effects on nucleic acids. , 1995, Current opinion in structural biology.

[123]  M. Record,et al.  Salt-nucleic acid interactions. , 1995, Annual review of physical chemistry.

[124]  M. Record,et al.  Salt dependence of oligoion-polyion binding: a thermodynamic description based on preferential interaction coefficients , 1993 .

[125]  J A McCammon,et al.  Poisson-Boltzmann analysis of the lambda repressor-operator interaction. , 1992, Biophysical journal.

[126]  T. Cech,et al.  Defining the inside and outside of a catalytic RNA molecule. , 1989, Science.

[127]  W. Gilbert Origin of life: The RNA world , 1986, Nature.

[128]  Gerald S. Manning,et al.  Counterion binding in polyelectrolyte theory , 1979 .

[129]  H. Magdelenat,et al.  Study of the self‐diffusion coefficients of cations in the presence of an acidic polysaccharide , 1974, Biopolymers.

[130]  C. DeLisi,et al.  Biological sciences: mRNA is expected to form stable secondary structures , 1974, Nature.

[131]  Gerald S. Manning,et al.  Limiting Laws and Counterion Condensation in Polyelectrolyte Solutions I. Colligative Properties , 1969 .

[132]  J. R. Fresco,et al.  Renaturation of transfer ribonucleic acids through site binding of magnesium. , 1966, Proceedings of the National Academy of Sciences of the United States of America.