THE CHEMISTRY OF THE NUCLEIC ACIDS AND NUCLEOPROTEINS By J. M. Gutianp, G. R. Barker, AND D. O. JorpDan Department of Chemistry, University College, Nottingham, England This contribution makes no attempt to be a complete compendium of the literature but is designed as a critical survey of the present posi- tion in a few main sections of this field. Considerable difficulties and delays have been encountered in obtaining copies of certain journals and any omissions on this score are regretted. NUCLEOPROTEINS AND NUCLEIC ACIDS Isolation of nucleoproteins——-Many methods for isolating nucleo- proteins involve a stage, either during the extraction or more generally in the precipitation process, which is relatively drastic and may pro- duce an alteration in the chemical and physical properties of the nucleoprotein ; thus the majority of preparations involve precipitation of the extracted material with hydrochloric or acetic acids. The ex- traction processes vary considerably and employ water (1 to 13), dilute alkaline solution (14 to 17), sodium chloride solution (18, 19, 20), or buffer solutions of pH values ranging from 4 to 11 (21 to 26), followed in each case by precipitation with acid. It has been suggested (27) that nucleoproteins prepared thus are of variable composition, the precipitated nucleic acid carrying with it varying quantities of loosely bound protein, and these methods are now considered unsatis- factory (28) in view of the possible rupture of the nucleic acid-protein bond during the acid precipitation. A more controlled extraction of liver nucleoprotein is that in which the tissue was treated with a solu- tion containing 0.03M sodium bicarbonate and 0.5M potassium chloride (29, 30) and the nucleoprotein precipitated by adjusting the solution to pH 4.2. Even these conditions may, however, be too drastic, and more recently attention has been concentrated on modi- fications of the original mild methods (1 to 11, 15), coupled with precipitation of the nucleoprotein with saturated ammonium sulphate (12, 31) or calcium chloride (6, 7, 32) solutions. Thus an early method (32) has been modified (33) whereby fresh pulped calf thy- mus is extracted with water at 5° C. for twenty-four to thirty-six hours, and after clarification of the extract the nucleoprotein is pre- 175 -_ NUCLEIC ACIDS AND NUCLEOPROTEINS 199 differentiation of the cell, and from which, for instance, the secretory granules of the pancreas are derived. An interesting aspect of the nucleic acids of bacterial cells has been revived recently. For some time it has been known that Gram-positive pneumococci can become Gram-negative, and this change was brought about by extracting the cells in neutral solution (20) and by an en- zyme, apparently identical with the pancreas enzyme which acts upon yeast ribonucleic acid (226). The material released into the solution during the former process contained pentose nucleic acid and a nucleo- protein (20, 227). A similar change has now been effected in yeast cells and Gram-positive bacteria by extraction with a solution of a bile salt (228), and it was also possible to restore the Gram-positive re- action by replacing the responsible material, an essential component of which appears to be the magnesium salt of a pentose nucleic acid ; other salts of nucleic acid could not be plated back in this way. In agreement with the previous workers, the stainable material could be progressively extracted, that part on the surface of the cell being re- moved first. INDUCED TRANSFORMATION OF PNEUMOCOCCAL TYPES Among micro-organisms, the most striking example of the repro- ducible and controllable induction of inheritable and specific altera- tions in cell structure and function is the transformation of specific types of pneumococcus. This type of change has been brought about both in vivo and in vitro, and analogous transformations have been carried out in the field of viruses (for references, see 62). Avery, MacLeod & McCarty (62) have now isolated from type III pneu- mococci a biologically active fraction which in exceedingly minute amounts is capable under appropriate conditions of inducing the trans- formation of unencapsulated R variants of pneumococcus type II into fully encapsulated S cells of type III. Other variants are not transformed in this way. Examination of the active extract indicated, :within the limits of the methods employed, that protein, unbound? lipoid, and serologically active polysaccharide were absent, and that it consisted principally, if not solely, of a sodium salt, in homogeneous viscous form, of a desoxypentose nucleic acid of molecular weight of the order of 500,000. It is possible, as the authors suggest, that the biological activity of the material is not an inherent property of the nucleic acid but is due to minute amounts of some other substance so intimately associated with it as to escape detection. If, however, as Co uty — JAN 16 BS Gels 200 GULLAND, BARKER, AND JORDAN the evidence strongly suggests, the transforming principle is a sodium salt of a desoxypentose nucleic acid, this type of polynucleotide must be regarded not merély as structurally important but as functionally active in determining the biochemical activities and specific character- istics of pneumococcal cells. This would appear to be the first occasion on which specific transformation has been experimentally induced in vitro by a chemically defined substance, and its implications are of the greatest importance in the fields of genetics, virology, and cancer research. LITERATURE CITED . Umsen, F., Abderhalden’s Handbuch der biologischen Arbeitsmethoden, 8, 2 (1922) . . HAMMARSTEN, O., Z. physiol. Chem., 19, 19-37 (1894) . Steupen, H., Z. physiol. Chem., $3, 539-44 (1907) . Wontcemwury, J., Z. physiol. Chem., 37, 475-83 (1903) . . Linrenrecp, L., Abderhalden’s Handbuch der biologischen Arbeitsmetho- den, 8, 8 (1922) 6. BANG, 1, Hofmeister’s Beitrage chem. Physiol. Path., 4, 115-38 (1903) 7. BANG, 1, Hofmeister’s Beitrige chem. Physiol. Path., 4, 362-77 (1903) 8 Bane, 1, Hofmeister’s Beitrage chem. Physiol. Path., 4, 331-61 (1903) 9. Gaucez, A., AND Jones, W., Abderhalden’s Handbuch der biologischen Arbeitsmethoden, 8, 9 (1922) 10. Jonzs, W., ano Wurrrze, G. H., Am. J. 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P., AND JenreTTE, W. V., J. Natl. Cancer Inst. 1, 77-90 (1940) i EDITORIAL COMMITTEE H. J. ALMQUIST D. R. HOAGLAND J. M. LUCK Cc. L. A. SCHMIDT H. A. SPOEHR | ANNUAL REVIEW OF BIOCHEMISTRY JAMES MURRAY LUCK, Editor ‘ Stanford University JAMES H.C. SMITH, Associate Editor Carnegie Institution of Washington Division of Plant Biology Stanford University, California VOLUME XIV 1945 ANNUAL REVIEWS, INC. STANFORD UNIVERSITY P.O., CALIFORNIA | PTT COLIN MUNRO MacLEOD 213 11. C. M. MacLeod and M.R. Krauss, “Stepwise Intra-Vype Transformation of Pneumococcus from R to 8 by Way of a Various Intermediate in Capsular Polysac- charide Production.” Journal of Experimental Medicine, 86(1947);439-53;, ©. M,. Macleod and M. R. Krauss, “Transformation Reactions with Two Non-Allelic R Murants of the Same Strain of Pheumococcus PFype VILL” Journal of Experimental Medicine, 103(1956):623-38, 12. R. Austrian and C. M. MacLeod, “Acquisition of M Protein by Pheumococei through Transtormation Reactions.” Journal of Experimental Medicine, 89(U449): 451-60. 13. E. Otolenghi and C. M. Macleod, “Genetic Transformation among Living Pneumococci in the Mouse,” Proceedings of the National Academy of Sciences of the United States of America, 50(\963):417. 14. T.S. Kuhn, The Structure of Sctentific Revolutions, 2d ed. (Chicago: University of Chicago Press, 1970), pp. 93-94, 15. J. D. Watson, The Double Helix (New York: Atheneum, 1968). 16. Stent, “Prematurity and Uniqueness in Scientific Discovery.” 17. J. Lederberg, “Reply to H. V. Wyatt,” Nature, 289, no. 5369(1972):234. Mue! A Review of Biuchemistry, 147733-47. 19. Dubos, The Professor, the Institute, and DNA, 20. Lederberg, “Reply to H. V. Wyatt.” 21. bid. 22. Dubos, The Professor, the Institute, and DNA, p. 148, 23. J. Lederberg. 1959 Nobel Prize Acceptance Lecture, Royal Caroline Medico- Surgical Institute, Stockholm, 29 May 1959. 24. R. D. Hotchkiss, “The Genetic Chemistry of the Pneumococcal Transforma- tions.” Harvey Lecture, 24 January 1954, 25. A. D. Hershey and M. Chase, “Independent Function of Viral Proteins and Nucleic Acid in Growth of Bacteriophage,” Journal of General Physiology, 36(1951):39. 26. Textbook, Elementary, An Introduction to Human Genetics, ed. H. Eldon Sutton (New York: Hol, Rinehart and Winston, 1965), p. 70. 27. A syndrome principally produced by mycoplasma. 28. C. M. MacLeod, “Primary Atypical Pneumonia,” Medical Clinics of North America, 27(1943):670-86. 29. C. M. MacLeod, R. G. Hodges, M. Heidelberger, and W. G. Bernhard, “Pre- vention of Pneumococcal Pneumonia by Immunization with Specific Capsular Poly- saccharides,” Journal of Experimental Medicine, 82(1945):445-65. “The Chemistry and Metabolism of Bacteria,"[Annual | - 30. °R. Austrian, R-M. Douglas, G. Schiffman, ét al. “Prevention of Paeumo- coccal Pneumonia by Vaccination,” Transactions of the Association of American Physi- Gans, 89(1976): 184. 31. CM. MacLeod, “Chemotherapy of Pheumococcic Pneumonia,” Journal of the American Medical Association, 113( 1940): 1405. 32. MacLeod and Krauss, “Stepwise Intra- Type Transformation of Pneumococ- cus from R to S by Way of a Various Intermediate in Capsular Polysaccharide Production”; “Transformation Reactions with Two Non-Allelic R Mutants of the Same Strain of Pneumococcus Type VU." 33. Austrian and MacLeod. “Acquisition of M Protein by Pneumococci through Transformation Reactions.”