Types Of Base Pairs Research Articles

X-Ray Structural Studies of Adenosine 5′-Triphosphate Metal Compounds

Abstract Solid State structures of the ATP-metal compounds are critically analyzed. Particular attention is devoted to the metal coordination and its influence on the conformation of the triphosphate chain and ribose ring. The three phosphate groups are involved in the coordination in all the compounds. The strongest coordinating group is γ-phosphate, while α-phosphate is the weakest. The γ-phosphate can triphosphate chain reveals a high degree of flexibility about its backbone bond and torsion angles; however, it is always in a bent conformation with the P…P…P angle ranging from 85–100°. The O(βγ)-P(γ) bond is the longest of the backbone O–P linkages. The conformation of the ribose ring is commonly C(2′)- or C(3′)-endo; however, one ATP molecule has a C(4′)-endo geometry. The orientation of the adenine system about the glycosidic linkage is anti. The adenine system does not show any donating ability in most of the structures. Nevertheless, formatioin of a macrochelate through the coordination of adenine N(7) and γ-phosphate oxygen atoms to sodium ions was found in the structures of ATP-disodium salt. Furthermore, the adenine system is involved in stacking interactions and some type of base pairing via H-bridges in all the ATP-metal compounds. The presence of an aromatic base facilitates the formation of crystalline material. The base can act as a ligand for the metal cation, or it can just stabilize the crystal lattice via weak interactions such as stacking forces and H-bonds. Previously unpublished data for [Co(HATP)2][Co(H2O)6]·2(HDPA)·9H2O are reported. The review also includes data relevant to ADP-metal compounds.

Effect of supercoiling on the melting characteristics of heteropolynucleotides

Considering a supercoiled DNA molecule, having equal numbers of two distinct types of base-pairs, it has been shown theoretically that even for the extreme cases of mixing of the two types of base-pairs in a supercoiled DNA, the melting temperatures as well as the melting curves do not differ significantly. This indicates that these properties are practically independent of the detailed base sequence when the molecule is a covalently closed one and may be replaced by an equivalent homopolynucleotide whose binding energy is equal to the average base-pairing energy of the original DNA. This conclusion has been further supported by comparing the theoretical results with those obtained experimentally in the cases of polyoma DNA and phi X174 DNA. Finally, the effects of supercoiling on the cooperativity of melting and a few aspects of the differential melting characteristics of a supercoiled DNA have been discussed which provide a clear physical understanding of the process.

The Difference in the Type of Codon-Anticodon Base Pairing at the Ribosomal P-Site Is One of the Determinants of the Translational Rate1

By utilizing an enzymatically reconstructed tRNA variant containing an altered anticodon sequence, we have examined the different biochemical behavior of translation between the Watson-Crick type and the wobble type base pair interactions at the first anticodon position. We have found that the Watson-Crick type base pair has an advantage in translation in contrast to the wobble type base pair by comparing the efficiency of transpeptidation of native tRNA(Phe) (anticodon; GmAA) with its variant tRNA (anticodon; AAA) in the poly(U)-programmed ribosome system. Thomas et al. [Proc. Natl. Acad. Sci. U.S. (1988) 85, 4242-4246] showed that the wobble codon at the ribosomal A-site accepted its cognate tRNA less efficiently than the Watson-Crick base pairing codon. We report here that the wobble interaction at the ribosomal P-site also affected the rate of translation. This variable translational rate may be a mechanism of gene regulation through preferential codon usage.

Parallel Double Helices of DNA. Conformational Analysis of Regular Helices with the Second Order Symmetry Axis

We have performed a conformational analysis of DNA double helices with parallel directed backbone strands connected with the second order symmetry axis being at the same time the helix axis. The calculations were made for homopolymers poly(dA).poly(dA), poly(dC).poly(dC), poly(dG) poly(dG), and poly(dT).poly(dT). All possible variants of hydrogen bonding of base pairs of the same name were studied for each polymer. The maps of backbone chain geometrical existence were constructed. Conformational and helical parameters corresponding to local minima of conformational energy of "parallel" DNA helices, calculated at atom-atom approximation, were determined. The dependence of conformational energy on the base pair and on the hydrogen bond type was analysed. Two major conformational advantageous for "parallel" DNA's do not depend much on the hydrogen-bonded base pair type were indicated. One of them coincided with the conformational region typical for "antiparallel" DNA, in particular for the B-form DNA. Conformational energy of "parallel" DNA depends on the base pair type and for the most part is similar to the conformational energy of "antiparallel" B-DNA.

A.cntdot.T and C.cntdot.C+ base pairs can form simultaneously in a novel multistranded DNA complex

Previous experiments have established that in certain synthetic oligomeric DNA sequences, including mixtures of d(AACC)5 with d(CCTT)5, adenine-thymine (A.T) base pairs form to the exclusion of neighboring protonated cytosine-cytosine (C.C+) base pairs [Edwards, E., Ratliff, R., & Gray, D. (1988) Biochemistry 27, 5166-5174]. In the present work, circular dichroism and other measurements were used to study DNA oligomers that represented two additional classes with respect to the formation of A.T and/or C.C+ base pairs. (1) One class included two sets of repeating pentameric DNA sequences, d(CCAAT)3-6 and d(AATCC)4,5. For both of these sets of oligomers, an increase in the magnitude of the long-wavelength positive CD band centered at about 280 nm occurred as the pH was lowered from 7 to 5 at 0.1 and 0.5 M Na+, indicating that C.C+ base pairs formed. Even though it may have been possible for these oligomers to form duplexes with two antiparallel A.T base pairs per pentamer, no A.T base pairing was detected by monitoring the CD changes at 250 nm. Thus, spectral data showed that as few as 40% C.C+ base pairs were stable in two sets of oligomers in which A.T base pairs did not form adjacent to, or in place of, C.C+ base pairs. (2) Another class of oligomer was represented by d(C4A4T4C4), which was studied by CD, HPLC, and centrifugation experiments. We confirmed previous work that this sequence was able to form both types of base pairs as the pH and temperature were lowered [Gray, D., Cui, T., & Ratliff, R. (1984) Nucleic Acids Res. 12, 7565-7580].(ABSTRACT TRUNCATED AT 250 WORDS)

Hairpin structures in DNA containing arabinofuranosylcytosine. A combination of nuclear magnetic resonance and molecular dynamics

Nuclear magnetic resonance (NMR) and model-building studies were carried out on the hairpin form of the octamer d(CGaCTAGCG) (aC = arabinofuranosylcytosine), referred to as the TA compound. The nonexchangeable protons of the TA compound were assigned by means of nuclear Overhauser effect spectroscopy (NOESY) and correlated spectroscopy (COSY). From a detailed analysis of the coupling data and of the NOESY spectra the following conclusions are reached: (i) The hairpin consists of a stem of three Watson-Crick type base pairs, and the two remaining residues, T(4) and dA(5), participate in a loop. (ii) All sugar rings show conformational flexibility although a strong preference for the S-type (C2'-endo) conformer is observed. (iii) The thymine does not stack upon the 3' side of the stem as expected, but swings into the minor groove. (This folding principle of the loop involves an unusual alpha t conformer in residue T(4).) (iv) At the 5'-3' loop-stem junction a stacking discontinuity occurs as a consequence of a sharp turn in that part of the backbone, caused by the unusual beta + and gamma t torsion angles in residue dG(6). (v) The A base slides over the 5' side of the stem to stack upon the aC(3) residue at the 3' side of the stem in an antiparallel fashion. On the basis of J couplings and a set of approximate proton-proton distances from NOE cross peaks, a model for the hairpin was constructed. This model was then refined by using an iterative relaxation matrix approach (IRMA) in combination with restrained molecular dynamics calculations. The resulting final model satisfactorily explains all the distance constraints.

Genetic alphabet for RNA gains two letters

Researchers in Switzerland have added two letters to the genetic alphabet, opening the possibility of synthesizing new types of RNA with greater catalytic potential. The genetic alphabet used by RNA molecules consists of only four letters, or bases: the purines adenine (A) and guanine (G), and the pyrimidines cytosine (C) and uracil (U). In double-stranded RNA, these bases, arranged along a sugar-phosphate backbone, hydrogen-bond to a complementary chain of bases so that each adenine is paired with a uracil and each guanine is paired with a cytosine. DNA differs from RNA only in that uracil is replaced by another pyrimidine, thymine (T), and the backbone sugar is deoxyribose instead of ribose. Thus, nature relies on only two types of base pairs (A-U or A-T, and G-C) to carry genetic information. But other types of base pairs are possible, as biological chemist Steven A. Benner and associates at the Swiss Federal Institute of Technology in Zurich have ...

Do Hoogsteen base pairs occur in DNA?

The importance of the Watson-Crick complementary base-pairing scheme has rather overshadowed alternative types of base pairs in DNA. One of these alternative base pairings, which is known as Hoogsteen pairing, is now receiving attention. Its presence in crystals of oligonucleotides bound to some antibiotics, and its possible existence in solution (and within long DNA fragments) remains to be unambiguously estimated. However, variability in DNA conformation appears to play an important biological role, and thus we should consider the presence of Hoogsteen base pairs as an interesting factor in inducing such changes.

Molecular characterization of two point mutants in the chloroplast atpB gene of the green alga Chlamydomonas reinhardtii defective in assembly of the ATP synthase complex

Two point mutants of Chlamydomonas reinhardtii, previously found by recombination and complementation analysis to map in the chloroplast atpB gene encoding the beta subunit of the CF1/CF0 ATP synthase, are here shown to be missense alterations near the 5' end of that gene. One mutant (ac-u-c-2-9) has a change at amino acid position 47 of the beta subunit from leucine (CTA) to arginine (CGA). In the second mutant (ac-u-c-2-29), the codon AAA (lysine) is changed to AAC (asparagine) at position 154. Spontaneous revertants of each mutant were isolated that restore the original wild type base pair. Northern analysis of total RNA and in vivo pulse labeling followed by immunoprecipitation reveals that both mutant atpB genes are transcribed and translated normally. However, immunoblots show that the amount of beta subunit associated with mutant thylakoids is only approximately 3% of that seen in wild type and that the CF1 alpha and gamma subunits are missing entirely. The disruption of ATP synthase complex assembly in these mutants is much more severe than in Escherichia coli beta subunit gene point mutants, which retain significant amounts of alpha and beta subunits on their membranes (Noumi, T., Oka, N., Kanazawa, H., and Futai, M. (1986) J. Biol. Chem. 261, 7070-7075). These results support the hypothesis that there are differences in assembly of the ATP synthase between E. coli and chloroplasts. In particular they indicate that beta must be present for assembly of the alpha and gamma subunits of CF1 onto chloroplast membranes.

DNA microstructural requirements for neocarzinostatin chromophore-induced direct strand cleavage

The microstructural requirements for optimal interaction of neocarzinostatin chromophore (NCS-C) with DNA have been investigated using a series of hexadeoxyribonucleotides with modified bases such as O6-methyl G (MeG), I, 5-methyl C (MeC), U, or 5-Bromo U (BrU) at specific sites in its preferred trinucleotide 5'GNaNb3':5'Na,Nb,C3' (Na = A, C, or T). Results show that MeG:C and G:MeC in place of G:C improve direct strand cleavage at the target Nb (Nb = T greater than A much greater than C greater than G), whereas MeC:G and C:MeG in place of Na:Nb, hinder cleavage. The optimal base target at Nb appears to be determined by its ability to form T:A type base pairing instead of C:G type. The observed differences in DNA strand cleavage patterns can be rationalized by induced changes in target site structure and are compatible with a model for NCS-C:DNA interaction in which the naphthoate moiety intercalates between 5'GNa3', and the activated tetrahydro-s-indacene, lying in the minor groove, abstracts a hydrogen atom from C-5' of Nb.

The 6-thioguanine/5-methyl-2-pyrimidinone base pair.

As part of a program to determine the physical possibility of expanding the number of types of base pairs in DNA, the pairing stabilities of the analog bases 6-thioguanine (GS) and 5-methyl-2-pyrimidinone (TH) in oligodeoxynucleotides were measured. Procedures were developed to synthesize oligodeoxynucleotides with the analog bases. The sequences of the synthesized oligomers were T-C-G-A-C-G-G-X-Y-C-C-G. An enzymatic procedure was developed to measure relative association constants of oligomer pairs with the self complementary reference oligomer, X = A and Y = T, K(T/A) = K. The results were K(C/G) = (5 +/- .5)K, K(TH/GS) = K/(1 +/- .5), K(T/G) = K/(9 +/- 3), K(TH/G) = K/(25 +/- 5), K(C/GS) less than K/30, K(TH/A) less than K/40, K(T/GS) less than K/40, K(C/A) less than K/40. The results with the standard bases are consistent with other methods of measurement. The stability of the base pair GS/TH is approximately the same as the standard base pair A/T.

Synthesis and proton-NMR studies of oligonucleotides containing an apurinic (AP) site.

In order to elucidate the conformational properties of base-deleted oligodeoxyribonucleotides, the molecules d-CpS(pCpG)n (n = 1,2; S = sugar) were synthesized by the phosphotriester method and characterized by 1H-NMR spectroscopy. Complete assignment of all non-exchangeable proton resonances of both compounds was obtained by 1D- and 2D-NMR techniques. In combination with computer simulation, these spectra yielded proton-proton and proton-phosphorus coupling constants of high accuracy. These data provide valuable information about the sugar and the backbone conformation. It appears that d-Cp1Sp2Cp3G4 does not form a duplex under any of the conditions studied. On the contrary, the base-deleted hexamer d-Cp1Sp2Cp3Gp4Cp5G6 occurs as a right-handed' staggered' DNA duplex at 280 K: the core of this duplex is formed by the residues C(3)-G(6); two 'dangling' residues C(1) and S(2) are located at the two 5'-ends of the duplex. The assignment of the corresponding imino proton resonances for [d-CpS(pCpG)2]2 was based on their thermal behavior: the line broadening of these resonances was studied as a function of temperature. The chemical shift and the number of imino proton resonances accord well with the number and type of Watson-Crick base pairs which can be formed in the staggered duplex described above. Thermodynamic parameters of duplex formation were obtained from an analysis of the chemical shift versus temperature profiles of aromatic base and H-1' protons. It is suggested that the cytosine ring of C(1) stacks, at least part of the time, with the guanine ring on the nucleotide residue, G(6), situated in the complementary strand. The binding of Lys-Trp-Lys to [d-CpS(pCpG)2]2 as well as to [d-CpGpCpG]1 was investigated. It is concluded that the indole ring of the tryptophan residue probably stacks on top of the 3'-terminal guanine base of both duplexes, but not on the nucleic acid bases next to the apurinic (AP) site.

In vivo D-serine deaminase transcription start sites in wild-type Escherichia coli and in dsdA promoter mutants.

The D-serine deaminase structural (dsdA) and regulatory (dsdC) genes are transcribed with opposite polarity from an intergenic region comprising more than 600 base pairs. The order of genes in the dsd region is supN-dsdA-dsdC-aroC---his. The DNA sequence of the intergenic region has been slightly revised from a previously published version (E. McFall and L. Runkel, J. Bacteriol. 154:1508-1512, 1983). The dsdA gene is preceded by a long open reading frame. The dsdA in vivo transcription start sites for the wild type (base pair +1) and for three phenotypically distinct promoter constitutive mutants were determined by the S1 nuclease method. They are identical and are located about 81 base pairs upstream of the translation start site. D-Serine deaminase regulation is normal in rho mutants. Possible mechanisms for dsdA activation are discussed.

Imino proton exchange in the 5S RNA of Escherichia coli and its complex with protein L25 at 490 MHz.

Imino proton exchange has been examined by NMR in the 5S RNA of Escherichia coli, its principal RNase A resistant fragment, fragment 1 (bases 1-11, 69-120), and complexes between that fragment and ribosomal protein L25 by using both real-time and relaxation techniques. Fragment 1 RNA imino protons exchange at rates between 0.5 and 15 s-1 at 303 K in 5 mM cacodylate buffer, pH 7.4. In contrast with many tRNAs, intact 5S RNA contains no imino protons with exchange lifetimes as great as 1 min. Consistent with the results of Gueron and his colleagues [Leroy, J. L., Bolo, N., Figueroa, N., Plateau, P., & Gueron, M. (1985) J. Biomol. Struct. Dyn. 2,915-939; Leroy, J. L., Broseta, D., & Gueron, M. (1985) J. Mol. Biol. 184, 165-178] with tRNA, exchange in 5S RNA is catalyst-limited under conditions generally used for imino proton spectroscopy, such as those given above. Using Gueron's catalyst saturation technique, base pair opening rates have been measured for several AU and GU base pairs in fragment 1. They range from 50 to 300 s-1 at 303 K and depend on base pair type and also to some degree on context. Similar studies have been done on complexes of L25 and fragment 1. The binding of L25 to fragment 1 reduces the exchange rate of many imino protons within the region to which it binds, consistent with the hypothesis that its binding stabilizes the secondary structure of 5S RNA.

Solution spectroscopy of dipicolinic acid interaction with nucleic acids: Role in spore heat resistance

The effect of dipicolinic acid (DPA) or its calcium chelate (CaDPA) on the spectral characteristics of nucleic acids was examined. Dipicolinic acid was found to displace ethidium gromide from DNA; this indicates that it may bind by intercalation. On interaction with DNA, the ultraviolet absorption spectrum revealed downfield shifts and caused progressive diminution in both DNA and dipicolinate chromophores. The strength and type of interaction may be ion-specific but not discriminatory to any type of base pairing. Spectral analysis also indicated that both dipicolinate and calcium dipicolinate bound to different RNA species, although the mechanism of binding was not elucidated. We conclude that the interaction of dipicolinate/ calcium dipicolinate with nucleic acids is a mechanism whereby water can be removed from spore polynucleotides, increasing their stability to denaturation.

Hairpin and duplex formation of the DNA octamer d(m5C-G-m5C-G-T-G-m5C-G) in solution. An NMR study.

The partly self-complementary DNA octamer d(m5C-G-m5C-G-T-G-m5C-G) was investigated by NMR spectroscopy in solution. It is demonstrated that this peculiar DNA fragment, under suitable conditions of concentration, salt and temperature, exclusively prefers to adopt a monomeric hairpin form with a stem of three Watson-Crick type base pairs and a loop of two residues. At high single strand concentration (8 mM DNA) and low temperature (i.e. below 295 K) the hairpin occurs in slow equilibrium with a B-dimer structure. At high ionic strength (greater than or equal to 100 mM Na+) and/or in the presence of methanol a third species appears, which is assigned to a Z-like dimer. In the B form, as well as in the Z dimer, the two central base pairs form G.T wobble pairs with the bases as major tautomers.

Hydrogen bond association constants of some nucleoside base pairs formed by an adenosine derivative and anticancer agents

Abstract Base-pair and trimer formation between certain adenosine derivatives and anticancer agents has been studied by Fourier-transform infrared spectroscopy by determining hydrogen-bond association constants. The results indicate that the anticancer agents studied have a preference for Hoogsteen over Watson-Crick type base-pair formation and that they have an appreciable tendency for trimer formation. This might be the way in which they interfere with site recognition and hinder unwarranted protein formation.

Expression of hepatitis B surface antigen with a recombinant adenovirus.

Early region 1 of the adenovirus type 5 genome was replaced with a DNA sequence containing the gene coding for the hepatitis B surface antigen (HBsAg) flanked by the major late promoter from adenovirus 2 and processing and polyadenylylation signals from simian virus 40. In one type of hybrid virus only the adenovirus 2 major late promoter, including just 33 base pairs of the adenovirus type 2 tripartite leader, preceded the coding region of the HBsAg gene. In another, this region was preceded by both the adenovirus major late promoter and almost the entire tripartite leader. The structure of the substituted sequence in each of the recombinant viral DNAs was identical to that in the plasmids used to construct the viruses. Approximately equivalent amounts of HBsAg-specific mRNA were produced late in infection with each recombinant virus. Although HBsAg production was detected late in infection of the hybrid virus not containing the full tripartite leader sequence, its level was 1/70th of that obtained with the hybrid virus containing this sequence. One likely interpretation is that the presence of the tripartite leader at the 5' end of this mRNA is critical for the synthesis of HBsAg polypeptide in the late stage of infection. HBsAg produced upon infection with the hybrid adenoviruses was glycosylated and secreted into the culture medium as particles that were essentially indistinguishable from the 22-nm particles found in human serum.

New light on tautomerism of purines and pyrimidines and its biological and genetic implications

The tautomerism of the natural 1-substituted pyrimidines and 9-substituted purines found in nucleic acids has been re-examined in the light of new experimental data on various nitrogen heterocycles in solution, in the gas phase and, in part, in low-temperature inert matrices. The results are compared with those obtained by quantum chemical calculations, including improved versions of the latter. Examples are presented of natural nucleosides which exhibit appreciable tautomerism in solution,e.g. formycins A and B, isoguanosine, but are not found in DNA. Illustrations are given of synthetic promutagenic nucleosides with pronounced tautomerism in solution relevant to their role in mutagenesis, such as the N4-hydroxy-and N4-methoxy cytidines. The amino-imino tautomeric equilibria of the promutagenic N6-hydroxy-and N6 -methoxy-adenosines are highly dependent on the solvent medium, the proportion of the imino species varying from 10% in CCl4 to 90% in aqueous medium. The type of base pairing of these is dependent on the conformation of the exocyclic hydroxy or methoxy groups. At the monomer level, addition of a potentially complementary base leads to a shift in the tautomeric equilibrium in favour of the species which pairs with this base. Biological and genetical implications of the foregoing are described.

Multistage unfolding of wheat germ ribosomal 5S RNA analyzed by differential scanning calorimetry.

Unfolding of purified wheat germ ribosomal RNA has been studied by differential scanning calorimetry (DSC) from 15 to 95 degrees C, in the presence and absence of 100 mM NaCl and/or 10 mM MgCl2. The total enthalpy of melting varies from about 290 (no sodium or magnesium) to 480 kcal/mol (Mg2+ only) and precisely accounts for the number and types of base pairs deduced from prior Fourier-transform infrared experiments. The composite DSC curves are analyzed into four individual two-state melting stages. Both Na+ and Mg2+ shift the melting transitions to higher temperature; in addition, Mg2+ causes the individual transitions to merge (i.e., melt cooperatively) and leads to irreversible chain cleavage at high temperature. The results are analyzed according to three general secondary base-pairing models for eukaryotic 5S RNA.