DNA G-quartets in a 1.86 A resolution structure of an Oxytricha nova telomeric protein-DNA complex.

The Oxytricha nova telomere end binding protein (OnTEBP) recognizes, binds and protects the single-stranded 3'-terminal DNA extension found at the ends of macronuclear chromosomes. The structure of this complex shows that the single strand GGGGTTTTGGGG DNA binds in a deep cleft between the two protein subunits of OnTEBP, adopting a non-helical and irregular conformation. In extending the resolution limit of this structure to 1.86 A, we were surprised to find a G-quartet linked dimer of the GGGGTTTTGGGG DNA also packing within the crystal lattice and interacting with the telomere end binding protein. The G-quartet DNA exhibits the same structure and topology as previously observed in solution by NMR with diagonally crossing d(TTTT) loops at either end of the four-stranded helix. Additionally, the crystal structure reveals clearly visible Na(+), and specific patterns of bound water molecules in the four non-equivalent grooves. Although the G-quartet:protein contact surfaces are modest and might simply represent crystal packing interactions, it is interesting to speculate that the two types of telomeric DNA-protein interactions observed here might both be important in telomere biology.

[1]  T. Cech,et al.  Effect of monovalent cation-induced telomeric DNA structure on the binding of Oxytricha telomeric protein. , 1990, Nucleic acids research.

[2]  T. Cech,et al.  Two versions of the gene encoding the 41-kilodalton subunit of the telomere binding protein of Oxytricha nova. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[3]  J. Feigon,et al.  Quadruplex structure of Oxytricha telomeric DNA oligonucleotides , 1992, Nature.

[4]  D. Rees,et al.  Conformational flexibility of B-DNA at 0.74 A resolution: d(CCAGTACTGG)(2). , 2000, Journal of molecular biology.

[5]  P. Kraulis A program to produce both detailed and schematic plots of protein structures , 1991 .

[6]  T. Cech,et al.  Cloning and expression of genes for the Oxytricha telomere-binding protein: Specific subunit interactions in the telomeric complex , 1991, Cell.

[7]  D. Gottschling,et al.  Telomere proteins: Specific recognition and protection of the natural termini of Oxytricha macronuclear DNA , 1986, Cell.

[8]  J. Feigon,et al.  Binding sites and dynamics of ammonium ions in a telomere repeat DNA quadruplex. , 1999, Journal of molecular biology.

[9]  A. Rich,et al.  Crystal structure of four-stranded Oxytricha telomeric DNA , 1992, Nature.

[10]  H M Berman,et al.  Hydration of the DNA bases is local. , 1995, Biophysical journal.

[11]  T. Cech,et al.  Properties of the telomeric DNA-binding protein from Oxytricha nova. , 1989, Biochemistry.

[12]  H M Berman,et al.  A systematic method for studying the spatial distribution of water molecules around nucleic acid bases. , 1993, Biophysical journal.

[13]  J. Feigon,et al.  Refined solution structure of the dimeric quadruplex formed from the Oxytricha telomeric oligonucleotide d(GGGGTTTTGGGG). , 1994, Structure.

[14]  D. Prescott,et al.  Telomeres of polytene chromosomes in a ciliated protozoan terminate in duplex DNA loops. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  D. Lilley,et al.  The crystal structure of a parallel-stranded guanine tetraplex at 0.95 A resolution. , 1997, Journal of molecular biology.

[16]  H. Berman,et al.  Hydration of the phosphate group in double-helical DNA. , 1998, Biophysical journal.

[17]  Aaron Klug,et al.  Telomeric DNA dimerizes by formation of guanine tetrads between hairpin loops , 1989, Nature.

[18]  J. Feigon,et al.  The effect of sodium, potassium and ammonium ions on the conformation of the dimeric quadruplex formed by the Oxytricha nova telomere repeat oligonucleotide d(G(4)T(4)G(4)). , 1999, Nucleic acids research.

[19]  R. Wellinger,et al.  The terminal DNA structure of mammalian chromosomes , 1997, The EMBO journal.

[20]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[21]  J. Griffith,et al.  Mammalian Telomeres End in a Large Duplex Loop , 1999, Cell.

[22]  D. Prescott,et al.  All gene-sized DNA molecules in four species of hypotrichs have the same terminal sequence and an unusual 3' terminus. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[23]  W. Gilbert,et al.  Formation of parallel four-stranded complexes by guanine-rich motifs in DNA and its implications for meiosis , 1988, Nature.

[24]  M. Horvath,et al.  Crystal Structure of the Oxytricha nova Telomere End Binding Protein Complexed with Single Strand DNA , 1998, Cell.

[25]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[26]  Z. Otwinowski,et al.  [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[27]  T. Cech,et al.  Telomeric protein-DNA point contacts identified by photo-cross-linking using 5-bromodeoxyuridine. , 1994, Biochemistry.

[28]  T. Cech,et al.  Telomeric DNA-protein interactions of Oxytricha macronuclear DNA. , 1987, Genes & development.

[29]  E. Blackburn,et al.  An overhanging 3' terminus is a conserved feature of telomeres , 1989, Molecular and cellular biology.

[30]  G. Thomas,et al.  A hairpin conformation for the 3' overhang of Oxytricha nova telomeric DNA. , 1998, Journal of molecular biology.

[31]  I. Brown,et al.  Empirical parameters for calculating cation–oxygen bond valences , 1976 .

[32]  J. Feigon,et al.  Strand orientation in the DNA quadruplex formed from the Oxytricha telomere repeat oligonucleotide d(G4T4G4) in solution. , 1993, Biochemistry.

[33]  G. Thomas,et al.  Molecular mechanism of DNA recognition by the alpha subunit of the Oxytricha telomere binding protein. , 1999, Biochemistry.

[34]  V A Zakian,et al.  Telomeres: Beginning to Understand the End , 1995, Science.

[35]  T. Cech,et al.  Monovalent cation-induced structure of telomeric DNA: The G-quartet model , 1989, Cell.

[36]  T. Cech,et al.  The β subunit of Oxytricha telomere-binding protein promotes G-quartet formation by telomeric DNA , 1993, Cell.

[37]  M Nayal,et al.  Valence screening of water in protein crystals reveals potential Na+ binding sites. , 1996, Journal of molecular biology.

[38]  P C Moody,et al.  The high-resolution crystal structure of a parallel-stranded guanine tetraplex. , 1994, Science.