On the Macromolecular Structure of Deoxyribonucleic Acid: An Interrupted Two-Strand Model

Dekker, Charles A.; Schachman, H. K.

doi:10.1073/pnas.40.10.894

Cited by 57 publications

(10 citation statements)

References 22 publications

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“…There is no evidence for the prior existence of interruptions in the chains to a degree greater than one in 3000. A slight preliminary attack by deoxyribo nuclease, which produces random breaks in the two chains (89,258,271), in the earlier study (89) could explain the discordance between the apparent molecular weights of heat-denatured DNA, which is now shown (99) to re tain its phosphodiester bonds intact. Although it has been shown that deoxy ribonuclease will break approximately 25 per cent of the phosphodiester bonds of DNA, liberating the bulk of the substrate as low molecular weight polynucleotides (di-to octanucleotides) (277), light-scattering measurements indicate an appa .…”

Section: Structure and Chemistrymentioning

confidence: 91%

“…The degradation of DNA.-The macromolecular changes in thymus DNA during the course of its degradation by acid, heat, and deoxyribonuclease were earlier summarized by Dekker & Schachman (89) and interpreted in terms of interrupted twin chains held together by hydrogen bonds. Re examination of these phenomena by Thomas (290), Doty (99,291), and Shumaker et al (271) more closely define this picture and establish that breaks in opposite chains, if separated by three to six hydrogen bonded base pairs, do not necessarily allow the dissociation of the resulting fragments.…”

Section: Structure and Chemistrymentioning

confidence: 99%

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The Nucleic Acids

Cohn¹,

Volkin²

1957

Annu. Rev. Biochem.

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Previous reviews of this topic have summarized progress in the biosyn thesis (43), preparation (5), an d organic chemistry of nucleic acids and nucleotides (37). This essay is concerned with recent work that deals with nucleic acid structure, relation to physiological and metabolic events, and biological interventions. The large increase in the number of published papers is met by an attempt to summarize rather than to expound'in detail. STRUCTURE AND CHEMISTRYThe ability to prepare DNA from various sources in a more purified and less degraded form and ever-increasing apparent molecular weight (100), coupled with an appreciation of the roles of hydrogen bonding and internal charge distribution, which underlie the structure proposed by Watson & Crick (305), have inspired re-examinations of the physicochemical charac teristics of DNA with improved methodology.Titration.-More accurate determination of the pC' values of the effec tive groups in herring sperm DNA by Cox et al. (71) indicates essentially no phosphoryl titration in the neutral range, which eliminates consideration of secondary phosphoryl end groups and thus the need to invoke triply esteri fied phosphate. The hysteresis (difference between forward and backward titrations) in the titration of both herring sperm (71) and thymus (48, 166) DNA is ascribed to the normal (A to T, C to G) hydrogen bonding, which is eliminated at the extremes of pH, in accordance with theory. The increase in ultraviolet absorption (hyperchromic effect) and bathochromic shift in the spectrum after DNA has been carried to pH 2 or 13 is also in accord (191). It has been indicated by Cavalieri & Rosenberg (46) that the ac ceptance of a proton by guanine, the apparent pK of which varies consider ably with ionic strength (46, 166), may cause rearrangements in the purine that result in breaking both guanine-cytosine hydrogen bonds. The same reaction is held accountable by Cavalieri et al. for a spectrophotometrically 1 The survey of the literature pertaining to this review was completed in Novem ber, 1956.2 The following standard abbreviations are employed: RNA and DNA for the ribo-and deoxyribonucleic acids; A, G, C, MC, D, and T for adenine, guanine, cytosine, methylcytosine, uracil, and thymine; AMP, ADP, etc., for the corresponding nucleoside 5'-phosphates and -pyrophosphates; and TMV for tobacco mosaic virus.• Our thanks are due to Dr. K. Sebesta, of the Czechoslovak Academy of Sciences, for critical sum�ries of pertinent papers appearing in some forty journals of Central European countries. 4 Manuscript prepared under Contract No. W-7405-eng-26 for the Atomic Energy Commission. 491 Annu. Rev. Biochem. 1957.26:491-522. Downloaded from www.annualreviews.org Access provided by University of Western Ontario on 02/07/15. For personal use only. Quick links to online content Further ANNUAL REVIEWS

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Section: Structure and Chemistrymentioning

confidence: 91%

Section: Structure and Chemistrymentioning

confidence: 99%

The Nucleic Acids

Cohn¹,

Volkin²

1957

Annu. Rev. Biochem.

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“…To address the topological problems and accommodate the results of ultracentrifugation data on DNA size, Stent’s Berkeley colleagues Howard Schachman and Charles Dekker proposed an alternative DNA model involving interrupted chains, which Stent found quite persuasive. Dekker & Schachman (1954); Holmes (2001), p. 45. …”

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confidence: 99%

Phosphorus-32 in the Phage Group: radioisotopes as historical tracers of molecular biology

Creager

2009

Studies in History and Philosophy of Science Part C: Studies in

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The recent historiography of molecular biology features key technologies, instruments and materials, which offer a different view of the field and its turning points than preceding intellectual and institutional histories. Radioisotopes, in this vein, became essential tools in postwar life science research, including molecular biology, and are here analyzed through their use in experiments on bacteriophage. Isotopes were especially well suited for studying the dynamics of chemical transformation over time, through metabolic pathways or life cycles. Scientists labeled phage with phosphorus-32 in order to trace the transfer of genetic material between parent and progeny in virus reproduction. Initial studies of this type did not resolve the mechanism of generational transfer but unexpectedly gave rise to a new style of molecular radiobiology based on the inactivation of phage by the radioactive decay of incorporated phosphorus-32. These ‘suicide experiments’, a preoccupation of phage researchers in the mid-1950s, reveal how molecular biologists interacted with the traditions and practices of radiation geneticists as well as those of biochemists as they were seeking to demarcate a new field. The routine use of radiolabels to visualize nucleic acids emerged as an enduring feature of molecular biological experimentation.

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“…The reactions were stopped and the products freed of unreacted deoxynucleoside triphosphate in one of two ways: (1) acidification with an equal volume of cold 1 N perchloric acid, then alternate solubilization of the precipitated product in 0.1 N NaOH and precipitation in 0.5 N perchloric acid (when enzyme had been omitted, no radioactivity could be detected in this repeatedly precipitated product); (2) addition of sodium chloride to a final concentration of 0.2 M and heating at 700 for 10 minutes,3 then repeated dialysis against 0.2 M sodium chloride. The radioactivity in this dialyzed product was then entirely precipitable by perchloric acid.…”

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confidence: 99%

Enzymatic Synthesis of Deoxyribonucleic Acid. Iv. Linkage of Single Deoxynucleotides to the Deoxynucleoside Ends of Deoxyribonucleic Acid

Adler¹,

Lehman²,

Bessman³

et al. 1958

Proc. Natl. Acad. Sci. U.S.A.

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Although the synthesis of DNA1 by an enzyme purified from Escherichia coli requires four deoxynucleoside triphosphates (dAPPP, dCPPP, dGPPP, TPPP) in addition to Mg++ and DNA,2 3 nevertheless a single deoxynucleoside triphosphate can be incorporated into DNA to an extent about one-thousandth of that for the complete reaction.3 The number of the single deoxynucleotides incorporated is of the same order as an approximation of the number of DNA chains added to the reaction mixture. It was the purpose of this investigation to determine whether the single deoxynucleotide, incorporated in the absence of the other three deoxynucleotide substrates, was covalently linked to the DNA and, if so, where in the DNA chain. It was also of interest to establish whether only one or several deoxynucleotides were added to a DNA chain. The results show that the single deoxynucleoside triphosphate reacts with the deoxynucleoside end of a DNA chain to form a typical 3',5'-phosphodiester linkage and that only one or a few nucleotides are added to a chain. METHODS AND MATERIALS Materials.-Uniformly labeled C'4-5'-deoxycytidylate was obtained by enzymatic hydrolysis of DNA from a Chromatium species grown on C1402 as the sole carbon source. Deoxyribonucleoside triphosphates were prepared as described previously.2 Micrococcal DNase4 was generously supplied by Dr. L. Cunningham. Bull semen 5'-nucleotidase5 and purified human semen phosphomonoesterase were kindly provided by Dr. L. A. Heppel. Calf spleen phosphodiesterase was prepared according to Hilmoe and Heppel.6 Venom phosphodiesterase was prepared from the venom of Crotalus adamanteus according to Koerner and Sinsheimer7 and was free of phosphomonoesterase. The DNA-synthesizing enzyme used in these studies (Fraction VII) was purified about 2,000-fold.2 Calf thymus DNA was prepared according to Kay, Simmons, and Dounce.8 Preparation of DNA products.-Four products were prepared from dAP32PP, dCP32PP, TP32PP, or Cl4dCPPP, essentially as described previously,2 except that the concentration of DNA was raised 25-fold in order to obtain greater amounts of incorporation. The reaction mixtures contained (per ml.) 6.0 mjumoles of a deoxynucleoside triphosphate; 6.7 ,umoles of magnesium chloride; 67 ,umoles of potassium phosphate buffer at pH 7.5; 800 jig. of calf thymus DNA; and 20 ,ug. of Fraction VII enzyme. The volume of these reaction mixtures ranged from 18 to 36 ml. Incubations were carried out at 370 for 2.5 hours (for dAP32PP, dCP32PP, and TP32PP) or for 1 hour (for C'4dCPPP).

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On the Macromolecular Structure of Deoxyribonucleic Acid: An Interrupted Two-Strand Model

Cited by 57 publications

References 22 publications

The Nucleic Acids

The Nucleic Acids

Phosphorus-32 in the Phage Group: radioisotopes as historical tracers of molecular biology

Enzymatic Synthesis of Deoxyribonucleic Acid. Iv. Linkage of Single Deoxynucleotides to the Deoxynucleoside Ends of Deoxyribonucleic Acid

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