Crystal Structure of an IHF-DNA Complex: A Protein-Induced DNA U-Turn

Integration host factor (IHF) is a small heterodimeric protein that specifically binds to DNA and functions as an architectural factor in many cellular processes in prokaryotes. Here, we report the crystal structure of IHF complexed with 35 bp of DNA. The DNA is wrapped around the protein and bent by >160 degrees, thus reversing the direction of the helix axis within a very short distance. Much of the bending occurs at two large kinks where the base stacking is interrupted by intercalation of a proline residue. IHF contacts the DNA exclusively via the phosphodiester backbone and the minor groove and relies heavily on indirect readout to recognize its binding sequence. One such readout involves a six-base A tract, providing evidence for the importance of a narrow minor groove.

[1]  R. Read Improved Fourier Coefficients for Maps Using Phases from Partial Structures with Errors , 1986 .

[2]  A M Gronenborn,et al.  Intercalation, DNA Kinking, and the Control of Transcription , 1996, Science.

[3]  Stephen K. Burley,et al.  1.9 Å resolution refined structure of TBP recognizing the minor groove of TATAAAAG , 1994, Nature Structural Biology.

[4]  M. Suzuki,et al.  Stereochemical basis of DNA bending by transcription factors. , 1995, Nucleic acids research.

[5]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[6]  R. Gumport,et al.  The isolation and characterization of mutants of the integration host factor (IHF) of Escherichia coli with altered, expanded DNA‐binding specificities. , 1992, The EMBO journal.

[7]  Helen M. Berman,et al.  Hydration of DNA , 1991 .

[8]  H. Berman Hydration of DNA: take 2 , 1994 .

[9]  Mike Carson,et al.  RIBBONS 2.0 , 1991 .

[10]  R. Gumport,et al.  Mutants of Escherichia coli integration host factor: DNA-binding and recombination properties. , 1994, Biochimie.

[11]  H. Nash,et al.  Comparison of protein binding to DNA in vivo and in vitro: defining an effective intracellular target. , 1995, The EMBO journal.

[12]  I. Tanaka,et al.  3-Å resolution structure of a protein with histone-like properties in prokaryotes , 1984, Nature.

[13]  A. Segall,et al.  Architectural elements in nucleoprotein complexes: interchangeability of specific and non‐specific DNA binding proteins. , 1994, The EMBO journal.

[14]  Sunghoon Kim,et al.  Mapping of a higher order protein-DNA complex: Two kinds of long-range interactions in λ attL , 1990, Cell.

[15]  H. Nash The HU and IHF Proteins: Accessory Factors for Complex Protein-DNA Assemblies , 1996 .

[16]  S. Nunes-Düby,et al.  Single base-pair precision and structural rigidity in a small IHF-induced DNA loop. , 1995, Journal of molecular biology.

[17]  H. Nash,et al.  The interaction of E. coli IHF protein with its specific binding sites , 1989, Cell.

[18]  N. Craig,et al.  E. coli integration host factor binds to specific sites in DNA , 1984, Cell.

[19]  T. Steitz,et al.  Crystal structure of a CAP-DNA complex: the DNA is bent by 90 degrees , 1991, Science.

[20]  W. McClure,et al.  Searching for and predicting the activity of sites for DNA binding proteins: compilation and analysis of the binding sites for Escherichia coli integration host factor (IHF). , 1990, Nucleic acids research.

[21]  T. Steitz,et al.  Crystal lattice packing is important in determining the bend of a DNA dodecamer containing an adenine tract. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Klug,et al.  The structure of an oligo(dA)·oligo(dT) tract and its biological implications , 1987, Nature.

[23]  Nora Goosen,et al.  The regulation of transcription initiation by integration host factor , 1995, Molecular microbiology.

[24]  I. Tanaka,et al.  A protein structural motif that bends DNA , 1989, Proteins.

[25]  J. Oberto,et al.  Serratia marcescens contains a heterodimeric HU protein like Escherichia coli and Salmonella typhimurium , 1996, Journal of bacteriology.

[26]  T. Steitz,et al.  Crystallization of Escherichia coli catabolite gene activator protein with its DNA binding site. The use of modular DNA. , 1990, Journal of molecular biology.

[27]  H. Nash,et al.  Specific photocrosslinking of DNA-protein complexes: identification of contacts between integration host factor and its target DNA. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[28]  G. Shaw,et al.  Anatomy of a Flexer–DNA Complex inside a Higher-Order Transposition Intermediate , 1996, Cell.

[29]  H. Berman,et al.  New parameters for the refinement of nucleic acid-containing structures. , 1996, Acta crystallographica. Section D, Biological crystallography.

[30]  D. Crothers Architectural elements in nucleoprotein complexes , 1993, Current Biology.

[31]  J. Molina-López,et al.  Geometry of the process of transcription activation at the sigma 54-dependent nifH promoter of Klebsiella pneumoniae. , 1994, The Journal of biological chemistry.

[32]  W. Messer,et al.  Functions of histone-like proteins in the initiation of DNA replication at oriC of Escherichia coli. , 1994, Biochimie.

[33]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[34]  P. van de Putte,et al.  Participation of the Flank Regions of the Integration Host Factor Protein in the Specificity and Stability of DNA Binding (*) , 1995, The Journal of Biological Chemistry.

[35]  R. Weisberg,et al.  Overproduction of Escherichia coli integration host factor, a protein with nonidentical subunits , 1987, Journal of bacteriology.

[36]  R. Lavery,et al.  Defining the structure of irregular nucleic acids: conventions and principles. , 1989, Journal of biomolecular structure & dynamics.

[37]  G. W. Hatfield,et al.  Transcriptional activation by protein-induced DNA bending: evidence for a DNA structural transmission model. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[38]  R. Gumport,et al.  Examining the contribution of a dA+dT element to the conformation of Escherichia coli integration host factor-DNA complexes. , 1996, Nucleic acids research.

[39]  J. Finch,et al.  Analysis of co-crystal structures to identify the stereochemical determinants of the orientation of TBP on the TATA box. , 1996, Nucleic acids research.

[40]  David A. Case,et al.  Structural basis for DNA bending by the architectural transcription factor LEF-1 , 1995, Nature.

[41]  S. Harrison,et al.  Effect of non-contacted bases on the affinity of 434 operator for 434 repressor and Cro , 1987, Nature.

[42]  R. Lavery,et al.  Measuring the geometry of DNA grooves , 1994, Biopolymers.

[43]  G J Kleywegt,et al.  xdlMAPMAN and xdlDATAMAN - programs for reformatting, analysis and manipulation of biomacromolecular electron-density maps and reflection data sets. , 1996, Acta crystallographica. Section D, Biological crystallography.

[44]  D. Goodsell,et al.  "...the tyranny of the lattice...". , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[45]  D. Lilley,et al.  DNA-protein: structural interactions , 1995 .

[46]  R. Gumport,et al.  The specific binding of Escherichia coli integration host factor involves both major and minor grooves of DNA. , 1995, Biochemistry.

[47]  Steven Hahn,et al.  Crystal structure of a yeast TBP/TATA-box complex , 1993, Nature.

[48]  H. Nash,et al.  Characterization of a set of integration host factor mutants deficient for DNA binding. , 1993, Journal of Molecular Biology.

[49]  T. Steitz,et al.  A DNA dodecamer containing an adenine tract crystallizes in a unique lattice and exhibits a new bend. , 1993, Journal of molecular biology.

[50]  R. Kaptein,et al.  Solution structure of the HU protein from Bacillus stearothermophilus. , 1995, Journal of molecular biology.

[51]  A. Rich,et al.  Asymmetric lateral distribution of unshielded phosphate groups in nucleosomal DNA and its role in DNA bending. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[52]  R. Huber,et al.  Accurate Bond and Angle Parameters for X-ray Protein Structure Refinement , 1991 .

[53]  A. Gronenborn,et al.  Molecular basis of human 46X,Y sex reversal revealed from the three-dimensional solution structure of the human SRY-DNA complex , 1995, Cell.

[54]  A. Oppenheim,et al.  Genetic and biochemical analysis of the integration host factor of Escherichia coli. , 1993, Journal of molecular biology.

[55]  V. de Lorenzo,et al.  Co-regulation by bent DNA. Functional substitutions of the integration host factor site at sigma 54-dependent promoter Pu of the upper-TOL operon by intrinsically curved sequences. , 1994, The Journal of biological chemistry.

[56]  G. Kleywegt,et al.  Halloween ... Masks and Bones , 1994 .

[57]  R. Gumport,et al.  Determining the DNA sequence elements required for binding integration host factor to two different target sites , 1994, Journal of bacteriology.

[58]  L. J. Maher,et al.  DNA bending by asymmetric phosphate neutralization. , 1994, Science.

[59]  Stephen K. Burley,et al.  Co-crystal structure of TBP recognizing the minor groove of a TATA element , 1993, Nature.

[60]  P. Vignais,et al.  The IHF proteins of Rhodobacter capsulatus and Pseudomonas aeruginosa. , 1994, Biochimie.

[61]  S. Goodman,et al.  Deformation of DNA during site-specific recombination of bacteriophage lambda: replacement of IHF protein by HU protein or sequence-directed bends. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[62]  K. Zhang SQUASH - combining constraints for macromolecular phase refinement and extension. , 1993, Acta crystallographica. Section D, Biological crystallography.

[63]  B. Lee,et al.  The interpretation of protein structures: estimation of static accessibility. , 1971, Journal of molecular biology.