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November 18, 1974, Vol 230, No. 7 >
ARTICLE | November 18, 1974

Ribonucleic Acid: Title and subTitle BreakI. The Test-Tube Synthesis of a Viral Nucleic Acid II. The Development and Use of Molecular Hybridization FREE

Sol Spiegelman, PhD
[+] Author Affiliations

Reprint requests to Institute of Cancer Research, Columbia University, 99 Fort Washington Ave, New York, NY 10032 (Dr. Spiegelman).


JAMA. 1974;230(7):1036-1042. doi:10.1001/jama.1974.03240070068041
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References

Two sets of investigations are briefly described. The first is aimed at an understanding of the molecular life history of lytic RNA virus with particular emphasis on the mechanism of RNA replication. The studies resulted in the isolation of the relevant RNA-directed RNA polymerase (replicase) and in the demonstration that this enzyme would mediate the continuous test-tube synthesis of unlimited quantities of infectious and biologically competent viral nucleic acid. This unique situation was exploited to examine the extracellular mutation and evolution of replicating molecules. The investigation culminated recently with the complete sequence determination of one of the mutant RNA molecules, an achievement that identified the secondary and tertiary structures as the source of the phenotypic wealth required to explain the evolutionary potential of these molecules.

The second investigation began with an attempt to provide a rigorous proof that DNA communicated its instructions via RNA complementary transcript. A procedure for annealing RNA to its complementary DNA and a sensitive method for detecting the resulting hybrid complexes were developed. Molecular hybridization thus provided the first direct evidence supporting the concept of cellular RNA as genetic messages. We further refined and simplified the procedure in an attempt to convert it into a powerful and flexible tool. RNA-DNA hybridization was then successfully used in the resolution of many of the central problems of molecular genetics. In this manner, nucleic acid hybridization became one of the most widely used molecular biologic techniques of the last decade. Its recent introduction into the cancer problem has provided the first molecular evidence of tumor viral information in human neoplasias.

(JAMA 230:1036-1042, 1974)

REFERENCES

1 +
Doi RH, Spiegelman S:  Homology test between the nucleic acid of an RNA virus and the DNA in the host cell . Science 138:1270-1272, 1962;.
2 +
Haruna I, Nozu K, Ohtaka Y, et al:  An RNA "replicase" induced by and selective for a viral RNA: Isolation and properties . Proc Natl Acad Sci USA 50:905-911, 1963;.
3 +
Haruna I, Spiegelman S:  Specific template requirements of RNA replicases . Proc Natl Acad Sci USA 54:579-587, 1965;.
4 +
Haruna I, Spiegelman S:  Autocatalytic synthesis of a viral RNA in vitro . Science 150:884-886, 1965;.
5 +
Spiegelman S, Haruna I, Holland IB, et al:  The synthesis of a self-propagating and infectious nucleic acid with a purified enzyme . Proc Natl Acad Sci USA 54:919-927, 1965;.
6 +
Pace NR, Spiegelman S:  In vitro synthesis of an infectious mutant RNA with a normal RNA replicase . Science 153:64-67, 1966;.
7 +
Spiegelman S, Haruna I, Pace NR, et al:  Studies in the replication of viral RNA, Gatlinburg Symposium . J Cell Physiol 70:35-64, 1967;.
8 +
Spiegelman S:  An in vitro analysis of a replicating molecule . Am Sci 55:221-265, 1967;.
9 +
Spiegelman S: Replication of viral RNA, Bertner Foundation Award Lecture, in Exploitable Molecular Mechanisms and Neoplasia . Baltimore, Williams & Wilkins Co, 1968;, pp 7-54.
10 +
Spiegelman S, Pace NR, Mills DR, et al:  The mechanism of RNA replication . Cold Spring Harbor Symp Quant Biol 33:101-124, 1968;.
11 +
Mills DR, Peterson RL, Spiegelman S:  An extracellular Darwinian experiment with a self-duplicating nucleic acid molecule . Proc Natl Acad Sci USA 58:217-224, 1967;.
12 +
Levisohn R, Spiegelman S:  The cloning of a self-replicating RNA molecule . Proc Natl Acad Sci USA 60:866-872, 1968;.
13 +
Levisohn R, Spiegelman S:  Further extracellular Darwinian experiments with replicating RNA molecules: Diverse variants isolated under different selective conditions . Proc Natl Acad Sci USA 63:805-811, 1969;.
14 +
Saffhill R, Schneider-Bernloehr H, Orgel LE, et al:  In vitro selection of bacteriophage Qβ ribonucleic acid variants resistant to ethidium bromide . J Mol Biol 51:531-539, 1970;.
15 +
Mills DR, Kramer FR, Spiegelman S:  Nucleotide sequence of a replicating RNA molecule with evolutionary potential . Science 180:916-928, 1973;.
16 +
Spiegelman S: On the nature of the enzyme-forming system, in Gaebler OH (ed): Enzymes , Henry Ford Hospital International Symposium. New York, Academic Press, 1956;, pp 67-89.
17 +
Spiegelman S, Kamen MD:  Genes and nucleoproteins in the synthesis of enzymes . Science 104:581-584, 1946;.
18 +
Spiegelman S: Nucleic acids and the synthesis of proteins, in McElroy WD, Glass HB (eds): The Chemical Basis of Heredity . Baltimore, The Johns Hopkins Press, 1957;, pp 232-267.
19 +
Spiegelman S: Protein and nucleic acid synthesis in subcellular fractions of bacterial cells, in Recent Progress in Microbiology , symposia, VII International Congress for Microbiology, 1958;, pp 81-103.
20 +
Spiegelman S:  Information transfer from the genome . Fed Proc 22:36-54, 1963;.
21 +
Spiegelman S, Hayashi M:  The present status of the transfer of genetic information and its control . Cold Spring Harbor Symp Quant Biol 28:161-181, 1963;.
22 +
Volkin E, Astrachan L:  Phosphorus incorporation of E coli RNA after infection with bacteriophage T2 . Virology 2:149-161, 1956;.
23 +
Nomura M, Hall BD, Spiegelman S:  Characterization of RNA synthesized in Escherichia coli after bacteriophage T2 infection . J Mol Biol 2:306-326, 1960;.
24 +
Doty P, Marmur J, Eigner J, et al:  Strand separation and specific recombination in deoxyribonucleic acids: Physical chemical studies . Proc Natl Acad Sci USA 46:461-471, 1960;.
25 +
Hall BD, Spiegelman S:  Sequence complementarity of T2-DNA and T2-specific RNA . Proc Natl Acad Sci USA 47:137-146, 1961;.
26 +
Gillespie D, Spiegelman S:  A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane . J Mol Biol 12:829-842, 1965;.
27 +
Yankovsky SA, Spiegelman S:  The identification of the ribosomal RNA cistron by sequence complementarity: I. Specificity of complex formation . Proc Nat Acad Sci USA 48:1069-1078, 1962;.
28 +
Yankovsky SA, Spiegelman S:  The identification of the ribosomal RNA cistron by sequence complementarity: II. Saturation of and competitive interaction at the RNA cistron . Proc Natl Acad Sci USA 48:1466-1472, 1962;.
29 +
Giacomoni D, Spiegelman S:  Origin and biologic individuality of the genetic dictionary . Science 138:1328-1331, 1962;.
30 +
Yankofsky SA, Spiegelman S:  Distinct cistrons for the two ribosomal RNA components . Proc Natl Acad Sci USA 49:538-544, 1963;.
31 +
Hayashi M, Spiegelman S:  The selective synthesis of informational RNA in bacteria . Proc Natl Acad Sci USA 47:1564-1580, 1961;.
32 +
Hayashi M, Spiegelman S, Franklin NC, et al:  Separation of the RNA message transcribed in response to a specific inducer . Proc Natl Acad Sci USA 49:729-736, 1963;.
33 +
Hayashi M, Hayashi MN, Spiegelman S:  Restriction of in vivo genetic transcription to one of the complementary strands of DNA . Proc Natl Acad Sci USA 50:664-672, 1963;.
34 +
Hayashi M, Hayashi MN, Spiegelman S:  DNA circularity and the mechanism of strand selection in the generation of genetic messages . Proc Natl Acad Sci USA 51:351-359, 1964;.
35 +
Ritossa FM, Spiegelman S:  Localization of DNA complementary to ribosomal RNA in the nucleolus organizer region of Drosophila melanogaster . Proc Natl Acad Sci USA 53:737-745, 1965;.
36 +
Ritossa FM, Atwood KC, Spiegelman S:  A molecular explanation of the bobbed mutants of Drosophila melanogaster as partial deficiencies of ribosomal DNA . Genetics 54:819-834, 1966;.
37 +
Spiegelman S, Burny A, Das MR, et al:  DNA-directed DNA polymerase activity in oncogenic RNA viruses . Nature 227:1029-1031, 1970;.
38 +
Spiegelman S, Burny A, Das MR, et al:  Synthetic DNA-RNA hybrids and RNA-RNA duplexes as templates for the DNA polymerases of the oncogenic RNA viruses . Nature 228:430-432, 1970;.
39 +
Axel R, Schlom J, Spiegelman S:  Presence in human breast cancer of RNA homologous to mouse mammary tumour virus RNA . Nature 235:32-36, 1972;.
40 +
Kufe D, Hehlmann R, Spiegelman S:  Human sarcomas contain RNA related to the RNA of a mouse leukemia virus . Science 175:182-185, 1972;.
41 +
Hehlmann R, Kufe D, Spiegelman S:  Viral-related RNA in Hodgkin's disease and other human lymphomas . Proc Natl Acad Sci USA 69:1727-1731, 1972;.
42 +
Axel R, Gulati SC, Spiegelman S:  Particles containing RNA-instructed DNA polymerase and virus-related RNA in human breast cancers . Proc Natl Acad Sci USA 69:3133-3137, 1972;.
43 +
Spiegelman S, Axel R, Baxt W, et al: Human cancer and the RNA tumor viruses, in RNA Viruses/Ribosomes . Amsterdam, North-Holland Publishing Co, 1972;, pp 73-99.
44 +
Kufe D, Hehlmann R, Spiegelman S:  RNA related to that of a murine leukemia virus in Burkitt's tumors and nasopharyngeal carcinomas . Proc Natl Acad Sci USA 70:5-9, 1973;.
45 +
Kufe D, Magrath IT, Ziegler JL, et al:  Burkitt's tumors contain particles encapsulating RNA-instructed DNA polymerase and high molecular weight virus-related RNA . Proc Natl Acad Sci USA 70:737-741, 1973;.

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Country-Specific Mortality and Growth Failure in Infancy and Yound Children and Association With Material Stature

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1 +
Doi RH, Spiegelman S:  Homology test between the nucleic acid of an RNA virus and the DNA in the host cell . Science 138:1270-1272, 1962;.
2 +
Haruna I, Nozu K, Ohtaka Y, et al:  An RNA "replicase" induced by and selective for a viral RNA: Isolation and properties . Proc Natl Acad Sci USA 50:905-911, 1963;.
3 +
Haruna I, Spiegelman S:  Specific template requirements of RNA replicases . Proc Natl Acad Sci USA 54:579-587, 1965;.
4 +
Haruna I, Spiegelman S:  Autocatalytic synthesis of a viral RNA in vitro . Science 150:884-886, 1965;.
5 +
Spiegelman S, Haruna I, Holland IB, et al:  The synthesis of a self-propagating and infectious nucleic acid with a purified enzyme . Proc Natl Acad Sci USA 54:919-927, 1965;.
6 +
Pace NR, Spiegelman S:  In vitro synthesis of an infectious mutant RNA with a normal RNA replicase . Science 153:64-67, 1966;.
7 +
Spiegelman S, Haruna I, Pace NR, et al:  Studies in the replication of viral RNA, Gatlinburg Symposium . J Cell Physiol 70:35-64, 1967;.
8 +
Spiegelman S:  An in vitro analysis of a replicating molecule . Am Sci 55:221-265, 1967;.
9 +
Spiegelman S: Replication of viral RNA, Bertner Foundation Award Lecture, in Exploitable Molecular Mechanisms and Neoplasia . Baltimore, Williams & Wilkins Co, 1968;, pp 7-54.
10 +
Spiegelman S, Pace NR, Mills DR, et al:  The mechanism of RNA replication . Cold Spring Harbor Symp Quant Biol 33:101-124, 1968;.
11 +
Mills DR, Peterson RL, Spiegelman S:  An extracellular Darwinian experiment with a self-duplicating nucleic acid molecule . Proc Natl Acad Sci USA 58:217-224, 1967;.
12 +
Levisohn R, Spiegelman S:  The cloning of a self-replicating RNA molecule . Proc Natl Acad Sci USA 60:866-872, 1968;.
13 +
Levisohn R, Spiegelman S:  Further extracellular Darwinian experiments with replicating RNA molecules: Diverse variants isolated under different selective conditions . Proc Natl Acad Sci USA 63:805-811, 1969;.
14 +
Saffhill R, Schneider-Bernloehr H, Orgel LE, et al:  In vitro selection of bacteriophage Qβ ribonucleic acid variants resistant to ethidium bromide . J Mol Biol 51:531-539, 1970;.
15 +
Mills DR, Kramer FR, Spiegelman S:  Nucleotide sequence of a replicating RNA molecule with evolutionary potential . Science 180:916-928, 1973;.
16 +
Spiegelman S: On the nature of the enzyme-forming system, in Gaebler OH (ed): Enzymes , Henry Ford Hospital International Symposium. New York, Academic Press, 1956;, pp 67-89.
17 +
Spiegelman S, Kamen MD:  Genes and nucleoproteins in the synthesis of enzymes . Science 104:581-584, 1946;.
18 +
Spiegelman S: Nucleic acids and the synthesis of proteins, in McElroy WD, Glass HB (eds): The Chemical Basis of Heredity . Baltimore, The Johns Hopkins Press, 1957;, pp 232-267.
19 +
Spiegelman S: Protein and nucleic acid synthesis in subcellular fractions of bacterial cells, in Recent Progress in Microbiology , symposia, VII International Congress for Microbiology, 1958;, pp 81-103.
20 +
Spiegelman S:  Information transfer from the genome . Fed Proc 22:36-54, 1963;.
21 +
Spiegelman S, Hayashi M:  The present status of the transfer of genetic information and its control . Cold Spring Harbor Symp Quant Biol 28:161-181, 1963;.
22 +
Volkin E, Astrachan L:  Phosphorus incorporation of E coli RNA after infection with bacteriophage T2 . Virology 2:149-161, 1956;.
23 +
Nomura M, Hall BD, Spiegelman S:  Characterization of RNA synthesized in Escherichia coli after bacteriophage T2 infection . J Mol Biol 2:306-326, 1960;.
24 +
Doty P, Marmur J, Eigner J, et al:  Strand separation and specific recombination in deoxyribonucleic acids: Physical chemical studies . Proc Natl Acad Sci USA 46:461-471, 1960;.
25 +
Hall BD, Spiegelman S:  Sequence complementarity of T2-DNA and T2-specific RNA . Proc Natl Acad Sci USA 47:137-146, 1961;.
26 +
Gillespie D, Spiegelman S:  A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane . J Mol Biol 12:829-842, 1965;.
27 +
Yankovsky SA, Spiegelman S:  The identification of the ribosomal RNA cistron by sequence complementarity: I. Specificity of complex formation . Proc Nat Acad Sci USA 48:1069-1078, 1962;.
28 +
Yankovsky SA, Spiegelman S:  The identification of the ribosomal RNA cistron by sequence complementarity: II. Saturation of and competitive interaction at the RNA cistron . Proc Natl Acad Sci USA 48:1466-1472, 1962;.
29 +
Giacomoni D, Spiegelman S:  Origin and biologic individuality of the genetic dictionary . Science 138:1328-1331, 1962;.
30 +
Yankofsky SA, Spiegelman S:  Distinct cistrons for the two ribosomal RNA components . Proc Natl Acad Sci USA 49:538-544, 1963;.
31 +
Hayashi M, Spiegelman S:  The selective synthesis of informational RNA in bacteria . Proc Natl Acad Sci USA 47:1564-1580, 1961;.
32 +
Hayashi M, Spiegelman S, Franklin NC, et al:  Separation of the RNA message transcribed in response to a specific inducer . Proc Natl Acad Sci USA 49:729-736, 1963;.
33 +
Hayashi M, Hayashi MN, Spiegelman S:  Restriction of in vivo genetic transcription to one of the complementary strands of DNA . Proc Natl Acad Sci USA 50:664-672, 1963;.
34 +
Hayashi M, Hayashi MN, Spiegelman S:  DNA circularity and the mechanism of strand selection in the generation of genetic messages . Proc Natl Acad Sci USA 51:351-359, 1964;.
35 +
Ritossa FM, Spiegelman S:  Localization of DNA complementary to ribosomal RNA in the nucleolus organizer region of Drosophila melanogaster . Proc Natl Acad Sci USA 53:737-745, 1965;.
36 +
Ritossa FM, Atwood KC, Spiegelman S:  A molecular explanation of the bobbed mutants of Drosophila melanogaster as partial deficiencies of ribosomal DNA . Genetics 54:819-834, 1966;.
37 +
Spiegelman S, Burny A, Das MR, et al:  DNA-directed DNA polymerase activity in oncogenic RNA viruses . Nature 227:1029-1031, 1970;.
38 +
Spiegelman S, Burny A, Das MR, et al:  Synthetic DNA-RNA hybrids and RNA-RNA duplexes as templates for the DNA polymerases of the oncogenic RNA viruses . Nature 228:430-432, 1970;.
39 +
Axel R, Schlom J, Spiegelman S:  Presence in human breast cancer of RNA homologous to mouse mammary tumour virus RNA . Nature 235:32-36, 1972;.
40 +
Kufe D, Hehlmann R, Spiegelman S:  Human sarcomas contain RNA related to the RNA of a mouse leukemia virus . Science 175:182-185, 1972;.
41 +
Hehlmann R, Kufe D, Spiegelman S:  Viral-related RNA in Hodgkin's disease and other human lymphomas . Proc Natl Acad Sci USA 69:1727-1731, 1972;.
42 +
Axel R, Gulati SC, Spiegelman S:  Particles containing RNA-instructed DNA polymerase and virus-related RNA in human breast cancers . Proc Natl Acad Sci USA 69:3133-3137, 1972;.
43 +
Spiegelman S, Axel R, Baxt W, et al: Human cancer and the RNA tumor viruses, in RNA Viruses/Ribosomes . Amsterdam, North-Holland Publishing Co, 1972;, pp 73-99.
44 +
Kufe D, Hehlmann R, Spiegelman S:  RNA related to that of a murine leukemia virus in Burkitt's tumors and nasopharyngeal carcinomas . Proc Natl Acad Sci USA 70:5-9, 1973;.
45 +
Kufe D, Magrath IT, Ziegler JL, et al:  Burkitt's tumors contain particles encapsulating RNA-instructed DNA polymerase and high molecular weight virus-related RNA . Proc Natl Acad Sci USA 70:737-741, 1973;.

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