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Biographical Memoirs: Volume 79 HOWARD M. TEMIN December 10, 1934–February 9, 1994 BY BILL SUGDEN HOWARD TEMIN LOVED knowledge, its acquisition, and its sharing. He pursued research where the logic of his experiments led him, independently of the scientific community’s initial skepticism toward his findings. He applied his expertise to improve public health policy to minimize smoking and to maximize benefits from research on the human immunodeficiency virus (HIV). This recounting of Howard Temin’s scientific career reflects my appreciation of his work as a tumor virologist. We shared lunch on Tuesdays for 20 years and gradually grew to be friends. Howard classified himself as a virologist and taught his students to be virologists. He taught the prominent course on animal virology on the University of Wisconsin’s Madison campus for 30 years. (I filled in for him when I first came to the McArdle Laboratory for Cancer Research while he traveled to Stockholm to accept the Nobel Prize. Typically, he insisted that I give several lectures before he left so that he could gauge whether I could lecture adequately and to coach me. I passed his test and I teach that course today.) During the first week I was in McArdle, Howard invited me to have lunch with him, Paul Kaesberg, and Roland Rueckert. These lunches were squeezed in for one-half hour before the weekly seminar on tumor virology—a training

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Biographical Memoirs: Volume 79 ground for formal presentations by graduate students and postdoctoral fellows that Howard had organized. I learned much from and about Howard at those lunches. He usually determined the subjects to be discussed but wanted our contributions. He valued science enormously while savoring the peccadilloes of its practitioners. These lunches, our frequent discussions about our faculty colleagues in McArdle, and our shared participation on many graduate student committees led us from formal collegiality to informal friendship. The uncited interpretations and motivations I ascribe to him come from these times together. Many of us who grew to know Howard professionally valued him personally. Rayla Greenberg Temin warmly described his rich personal life.1 Here I shall outline the depth and breadth of his scientific contributions. It is these contributions, coupled with his commitment to reason in all facets of his professional life, that made Howard Temin a major force in biology during the latter half of the twentieth century. THE PROVIRUS HYPOTHESIS Quite early in life Howard Temin was a devotee of science in general and biology in particular. He published his first paper at the age of 18 in 1953. He began the research he would follow professionally as a graduate student with Renato Dulbecco in Cal Tech in 1957. Not long before, Dulbecco and Margarite Vogt2 had developed a method to plaque poliomyelitis virus in cell culture; that is, they learned how to enumerate infectious virus particles in a stock of virus by detecting one focus of dead cells per infectious particle. The infected cells were lysed by expanding rounds of infection, viral replication, and cell death. This assay was the foundation for quantitative studies of lytic viruses in cell culture. As Howard began work in Dulbecco’s group, Mannaker and Groupé3 described an assay in which Rous

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Biographical Memoirs: Volume 79 sarcoma virus (RSV) altered the morphology of cells infected in culture. Here each focus of infection did not result from ensuing rounds of cell death but rather in survival of cells with altered phenotypes of shape. This in vitro assay was exciting because it could reflect in vitro the known ability of RSV to cause cancers in vivo soon after its inoculation into newborn chicks. Howard, working with Harry Rubin, a postdoctoral fellow in Dulbecco’s group, refined this assay and used it to investigate “morphological transformation” of cells by RSV in culture. Temin and Rubin (1958) refined the assay for RSV by overlaying chick embryo fibroblast (CEF) cells with agar soon after their exposure to dilutions of a virus stock. This overlay minimized the spread of progeny virus from an initially infected cell to distant cells, thereby confining the progeny from initial infections each to a single focus. The agar also helped to restrain the infected cells, which became less adherent to their initial site. Overlaid, infected cells yielded foci that enlarged exponentially with time, and this enlargement resulted primarily from division of the infected cells (1958). These foci arose linearly as a function of the dilution of the virus stock assayed over a large range of dilutions; these results indicated that a single particle of RSV is competent to initiate a focus (1958). Rubin and Temin (1959) used this quantitative focus assay for RSV to analyze its initiation of infection radiologically. They compared the sensitivities to exposure to X rays and ultraviolet (UV) light of RSV and New Castle disease virus (NDV) in initiating or maintaining infection. NDV infects CEF cells only lytically and has an RNA genome, as RSV was then thought to have. (Bather4 found RNA in semi-purified RSV. Crawford and Crawford5 developed an isopycnic method to purify RSV further and confirmed its genome as being RNA). They found that infection by NDV was resistant to

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Biographical Memoirs: Volume 79 previous exposure of the host cells to X rays or UV light, while infection by RSV was exquisitely sensitive to similar treatments. Once cells were infected with RSV, their ability to produce progeny virus had an intermediate sensitivity to treatment with X rays. The exquisite sensitivity of RSV to initiate infection of radiologically treated cells equaled that of treated CEF cells to form colonies. This finding indicated that initiation of infection by RSV shared a common radiosensitive target with cell division, in marked contrast to the radiologically resistant, cytocidal NDV. The reduced sensitivity to radiological treatment of RSV production after establishment of its infection, coupled with its exquisite sensitivity for initiation of infection, appeared similar to those of the temperate phage (1959). These studies led them to hypothesize that “the genome of the Rous sarcoma virus must be integrated with that of the cell before virus production can begin” (1959). This hypothesis drove much of Howard Temin’s research for the next 11 years. Temin and Rubin6 established the previously suspected model that cells in which RSV infection is established pass on to their progeny the capacity to release RSV. They analyzed single infected cells in microdrops and found both that the cells divided and that the daughter cells could release virus. This observation underscored the difference between RSV and lytic viruses such as EMC and NDV. RSV infection did not dramatically alter its host cell’s survival, but it did affect it genetically; lytic viruses merely killed their host cells. This realization provided another similarity to the temperate phage, and a difference. Temperate phage pass on phage genomes from infected parent to infected daughter cell; however, they do not continuously release progeny phage. In 1959 Harry Rubin moved to Berkeley, while Howard Temin continued his work at Cal Tech. Howard characterized

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Biographical Memoirs: Volume 79 isolates of RSV that conferred distinctive morphologies on their infected cells. He infected CEF cells that he had cloned to demonstrate that the distinctive cellular morphologies resulting upon infection with different isolates of RSV reflected genetic contributions of the viruses (1960). The capacity to endow an infected host cell with one morphology could mutate, the mutant virus then conferring on the cells it infects a new, distinctive morphology. Howard concluded that “the virus becomes equivalent to a cellular gene controlling cellular morphology.” He also contemplated the possibility that the ability of RSV to control the morphology of cells infected in vitro was related to the virus’s tumorigenic capacity in vivo. In 1960 Howard moved to the McArdle Laboratory for Cancer Research, where he carried on his research for the rest of his life. By this time an appreciation of the functions of cellular RNAs was crystallizing. Two groups published their findings, indicating that ribosomal RNAs were stable, structural elements of ribosomes, whereas short-lived RNAs conveyed information from DNA to the ribosomes to encode protein synthesis.7,8 This short-lived RNA is messenger RNA (mRNA). The other component of cellular RNA was transfer RNA (tRNA), small stable RNAs required to move amino acids to the ribosomes. The recognized functions of cellular RNAs did not include long-lived transfer of information. The means by which RSV, an RNA tumor virus, could stably affect the heritable morphology of infected cells was therefore enigmatic both for Howard and the scientific community at large. At the McArdle Laboratory Howard continued to study RSV infection genetically. He isolated and characterized cells infected with RSV arising after a low multiplicity of infection, which were morphologically altered but did not in general produce virus. Virus production could be rescued

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Biographical Memoirs: Volume 79 by infection of these converted non-virus-producing (CNVP) cells with a different strain of RSV. These CNVP cells induced tumors in susceptible chicks, as did virus-producing cells (1963,1). These experiments separated virus production from virus-mediated morphological transformation of the infected host cell and tumorigenicity. They demonstrated that RSV-infected cells could maintain the information for producing virus in the absence of such production. The information in the infected cell necessary to produce virus was designated the provirus. Howard analyzed the mechanism of infection by RSV biochemically. He assessed actinomycin D as an inhibitor of RSV infection and production. Actinomycin D had been recently shown to inhibit DNA-dependent RNA synthesis,9 but not the replication of some RNA viruses.10 At low concentrations (0.1 to 0.2 µg/ml), actinomycin reversibly inhibited infection by and production of RSV (1963,2). Concentrations up to 10 µg/ml had no effect on lytic infections by NDV (1963,2). Thus the effect of this inhibitor varied dramatically for the lytic RNA virus NDV and the transforming RNA tumor virus, RSV. Howard “suggested that the template responsible for synthesis of viral (RSV) nucleic acid either is DNA or is located on DNA” (1963,2). This unorthodox suggestion was a logical outgrowth of Temin’s genetic and biochemical studies of RSV replication in cell culture. Similar experiments demonstrating the sensitivity of RSV infection to treatment with actinomycin D were also reported by other groups.11 Temin tested his suggestion directly with two recently developed methods to detect specific RNA/DNA hybrids. He labeled RSV RNA with tritiated uridine by propagating infected cells in labeled medium, isolating released virus, and purifying its genomic RNA. This labeled RNA was then hybridized to cellular DNAs isolated from uninfected and infected cells. The results of

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Biographical Memoirs: Volume 79 these experiments indicated that more DNA in infected cells was detected by its hybridization to RSV RNA than DNA in uninfected cells (1964). The specific c.p.m. of labeled RNA hybridized to cellular DNAs were extremely low—too low to be compelling today. However, the small signals were consistent with the provirus of RSV being DNA, as Howard had hypothesized. Howard analyzed the effects of serum on the proliferation of CEF cells in culture and recognized that the removal of serum inhibited the cells’ proliferation.12 He used this insight to manipulate cultures of cells such that they were partially synchronized within their proliferative cycle. He infected these partially synchronized cells with RSV and found that the capacity of infected cells to support virus production required the infected cells to pass through mitosis (1967). He found this requirement for mitosis when cultures were partially synchronized by removal of serum, by treatment with excess thymidine, or by treatment with colchicine (1967). In all cases, RSV was produced from infected cells after they passed through mitosis. These experiments did not allow him to distinguish between a requirement for mitosis to form the provirus or to activate the provirus to allow production of progeny virus. Additional experiments with synchronized cells did support the hypothesis that the provirus was composed of DNA. Howard treated cells arrested largely in the G2 phase of the cell cycle with cytosine arabinoside, an inhibitor of DNA synthesis. This treatment inhibited formation of the provirus as reflected by a failure to produce progeny RSV; the inhibition was abrogated by simultaneous treatment with deoxycytidine (1967). This and related observations indicated that formation of the provirus required DNA synthesis, but that DNA synthesis need not occur during the S phase of the cell cycle. The accumulated findings from Howard’s experiments analyzing infection by RSV in

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Biographical Memoirs: Volume 79 cell culture were convincing to him, but still did not persuade virologists in general. The years 1967 and 1968 were watershed occasions for animal virology; at least three findings with DNA and RNA viruses would eventually contribute to Howard’s appreciation of the life cycle of RSV and to its general acceptance by virologists. Joe Kates and Brian McAuslan,13 working with rabbit poxvirus, a member of the same family of DNA viruses as is smallpox, demonstrated that purified viral core particles contain a DNA-dependent RNA polymerase. The activity was detected only after intact viral particles were disrupted, was inhibited by actinomycin D, and synthesized RNA homologous to rabbit poxviral DNA. (It has since been demonstrated that poxviral DNA-dependent RNA polymerase is encoded by the virus and related to similar cellular enzymes.) In 1968 Aaron Shatkin and J.D.Sipe identified an RNA-dependent RNA polymerase in cores of reoviruses.14 Reoviruses contain multiple distinct segments of double-stranded RNA within their inner core. Shatkin and Sipe found that they needed to remove the virus’s outer protein shell to detect the polymerase activity, which was dependent on the addition of all four ribonucleoside triphosphates and synthesized RNAs homologous to reoviral genomic RNA. Thus, by the end of 1968 it was evident that both a DNA-and an RNA-containing animal virus house a template-dependent RNA polymerase activity. Studies with the DNA tumor viruses, simian virus 40 (SV40) and polyoma (Py) in 1968 demonstrated that these tumor viruses affected their transformation of cells in culture by maintaining their DNA genomes integrated into their host cell’s chromosomes. Joe Sambrook—a postdoctoral fellow with Howard’s former mentor Renato Dulbecco—led a team of researchers who isolated chromosomal DNAs from SV40-and Py-transformed cells and showed by nucleic acid hybridi-

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Biographical Memoirs: Volume 79 zation that these cellular DNAs had integrated copies of the viral DNAs.15 Two approaches were critical to the success of their experiments. In one approach the isolated cellular DNAs were separated on alkaline sucrose gradients by velocity sedimentation, which minimized any fortuitous, non-covalent association of viral DNAs with large chromosomal DNAs. In all experiments the integrated viral DNAs were detected by hybridization with viral RNAs synthesized in vitro with DNA-dependent RNA polymerase isolated from E. coli. This latter approach permitted the radiolabeled RNA synthesized in vitro with purified viral DNA templates to be of high specific activity. The results from the hybridization experiments were thus compelling and demonstrated that certain DNA tumor viruses maintained their genomes as DNA integrated into the virally transformed host’s chromosomes. As these results with different animal viruses were being appreciated, David Boettiger began working as a graduate student at the McArdle Laboratory with Howard Temin. Colleagues at McArdle had been working previously with various halogenated nucleotides. Charlie Heidelberger had synthesized fluorodeoxyuracil (5-FU) and pioneered its use in chemotherapy for certain human cancers. Waclaw Szybalski had studied bromodeoxyuridine (5'-BUdR) and had shown that, on its incorporation into the DNA of bacteria, DNA became sensitized to damage induced by exposure of the cells to near ultraviolet or visible light. David, with Howard’s guidance, rendered CEF cells stationary by withdrawing serum from their medium, infected them with RSV, treated them with 5'-BUdR, and exposed them to near UV light (1970). The stationary cells did not incorporate 5'-BUdR and were not detectably harmed by the near UV light. However, this regimen reduced infection by RSV to 5 percent of that of the untreated control. Importantly, increasing the multiplicity of infection twenty-fold significantly decreased the

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Biographical Memoirs: Volume 79 rate of inactivation of infection, indicating that multiple virus particles were inactivated independently within a single infected cell. This experiment provided powerful support for Howard’s provirus hypothesis. It demonstrated that the incorporation of a light-sensitive DNA-nucleotide analogue soon after infection by RSV, followed by exposure to near UV light, inactivated that infection without damaging stationary host cells. This manuscript was submitted to Nature in March of 1970, but it was not immediately accepted. While David Boettiger was optimizing his inactivation experiments, Satoshi Mizutani, a postdoctoral fellow with Howard, pursued the possibility foreshadowed by the findings with rabbit poxvirus and reovirus that RSV might contain a polymerase activity capable of copying RSV RNA into a precursor to its proviral DNA. Permeabilization of the envelope of the viral particle with a non-ionic detergent in the presence of dithiothreitol allowed detection of the activity soon to be dubbed reverse transcriptase. This enzyme could incorporate the four deoxyribonucleotides to yield DNA. Treatment of the disrupted virions with RNase A prior to their incubation with labeled deoxynucleotides abrogated subsequent DNA synthesis, indicating that RSV contained an RNA-dependent DNA polymerase. Mizutani and Temin submitted their findings to Nature on June 15, 1970. Their paper was published on June 27, 1970 (1970,1). David Baltimore at MIT published similar findings for murine RNA tumor viruses in the same issue of Nature.16 He had submitted a manuscript in March to the Proceedings of the National Academy of Sciences relating his findings that vesicular stomatitis virus, VSV, a rhabdovirus, contained an RNA-dependent RNA polymerase, again focusing attention on the existence of template-dependent polymerases intrinsic to various families of viruses.17 The two reports of RNA-dependent DNA polymerase

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Biographical Memoirs: Volume 79 per replication cycle—extremely frequently. The recombinants were analyzed for loss of the diagnostic restriction endonuclease sites; most had lost them and represented genomic recombinants.60 Recombination within a retrovirus can be viewed to result from reverse transcriptase copying one template RNA or DNA and then switching to the other present in the viral particle. The high rate of recombination found by Hu and Temin indicates that such a template switch occurs once per every two or three cycles of retroviral replication. Hu and Temin built on their initial study by using two vectors, neo–/hyg+ and neo+/hyg–, which were marked throughout their genomes with eight different restriction endonuclease sites. Evidence from work on disparate retroviruses from many groups provided a detailed model for the synthesis of the linear precursor to a provirus that Hu and Temin outlined (1990, Figure 2). They generated and analyzed recombinants between the marked vectors and interpreted their formation in light of the detailed model for the synthesis of the proviral precursor. The first step in synthesis of the precursor to the provirus is synthesis of DNA primed from the tRNA near the 5' end of the viral RNA. This DNA shortly runs out of template and can be isolated from in vitro reactions as strong-stop DNA. Hu and Temin (1990) identified which endonuclease restriction sites were present in recombinant proviruses. They used these analyses to show that strong-stop DNA, which could in theory transfer to the 3' end of the RNA molecule from which it was synthesized or to the 3' end of the other RNA molecule within the virus particle, could in fact transfer to either template. Once the first strand of DNA was synthesized, they found that the second strand was primed only from the first strand’s template. Howard interpreted the high frequency of detected recombination during retroviral replication to result from

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Biographical Memoirs: Volume 79 the inherent ability of reverse transcriptase to switch its templates, an event essential to synthesis of the proviral precursor.61 Multiple studies with a variety of retroviral vectors and helper cells allowed Howard and his colleagues to prove the mechanism by which the provirus is synthesized. They now used this general approach to illuminate mechanisms by which retroviruses capture cellular proto-oncogenes to evolve into rapidly transforming derivatives. Jiayou Zhang developed helper cells with two non-homologous vectors. RNA synthesized from one would yield RU5 upstream of hygr but no U3 sequences. The other would yield RU5 upstream of neor along with U3R. Recombination within heterodimeric viruses released from these helper cells would be required to generate a functional 3' end of a hygr provirus (U3RU5 hygr U3RU5). Zhang and Temin (1993) infected cells with virus from characterized clones of helper cells with these proviruses and selected either for neor or hygr. Hygr cells arose at a frequency that indicated that nonhomologous recombination occurred at a rate of 0.1 percent to 1 percent of that of homologous recombination for retroviruses. They determined the sequence of the junctions of the recombinants and excitingly found a correlation with the kinds of junctions observed for sites at which retroviral sequences join viral oncogenes in rapidly transforming retroviruses.62 For example, in a group of non-homologous recombinants they described as being general, they found one example of recombination without sequence identity, six with a short region of five to eight base pairs of sequence identity, and three with insertions at the site of recombination. Inspection of naturally occurring, highly transforming retroviruses revealed that their sites of recombination usually contained short stretches of sequence identity or insertions.62 The non-homologous recombination they had characterized

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Biographical Memoirs: Volume 79 underlies the means by which retroviruses form their 3' ends while capturing cellular proto-oncogenes. HUMAN RETROVIRUSES Howard’s hypothesizing the provirus, his work on its structure and on its synthesis involved avian retroviruses. RSV and REV-T were known to cause sarcomas and lymphomas in chickens at a time when human retroviruses were not identified, let alone shown to cause human disease. Confirmation of the provirus hypothesis, however, focused attention on retroviruses and helped to justify the National Cancer Institute’s major investment in research in the 1970s on retroviruses in general and a search for human retroviruses in particular. Although he in some sense started this band, Howard did not follow its wagon. He continued to study avian retroviruses. By the end of the decade, Y.Hinuma, following a paradigm he developed to study Epstein-Barr virus, a human tumor virus in the herpes virus family, helped to identify human T-cell leukemia virus type 1 (HTLV-1), a bona fide leukemia-causing human retrovirus. Bob Gallo and his colleagues through their research on propagating human T cells in medium containing IL-2 also contributed to the identification of this human tumor virus. The 1980s subsequently brought the recognition of another pathogenic human retrovirus that would occupy Howard’s interest and eventually some of his research efforts. Acquired immunodeficiency syndrome (AIDS) was a disease entity identified in the early 1980s and found initially to be clustered in male homosexuals in the United States. The wasting that characterized AIDS was fatal, and suggestions for its cause varied widely and were often irrational. Luc Montagnier in the Institute Pasteur and Bob Gallo and his colleagues at the National Institutes of Health (NIH) identified a retrovirus linked to AIDS. Contentious claims

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Biographical Memoirs: Volume 79 of priority appeared for a while to cloud the importance of this discovery. Howard worked through an NIH committee to help name this virus and limit any contention—thus the neutrally named human immunodeficiency virus or HIV. Epidemiological studies provided increasingly robust data indicating that HIV causes AIDS, but a few virologists, such as Peter Duesberg, argued with this conclusion.63 Some of Howard’s colleagues suggested that such arguments be ignored, but he was very concerned that those arguments be addressed directly and refuted. In 1988 Howard joined Bill Blattner and Bob Gallo to publish an article entitled “HIV Causes AIDS” in Science (1988). Here he contributed not original research but rather his depth and breadth of reading as well as his scientific integrity to reason for the well-being of people. He repeated his reasoned address in a short article in Policy Review in 1990.64 Howard willingly involved himself in national policy decisions to promote the best possible research on AIDS. Between 1985 and 1994 Howard served on 12 national and international committees focused on multiple facets of HIV and AIDS. These included an oversight committee on AIDS activities of the Institute of Medicine from 1987 to 1990, chairing the HIV Genetic Variation Advisory Panel for the National Institute of Allergies and Infectious Diseases from 1988 to 1994, the Global Commission on AIDS from 1991 to 1992, and the World Health Organization Advisory Council on HIV and AIDS from 1993 to 1994. He devoted more of his efforts to guiding policies on AIDS and HIV than on any other public health problem faced during his professional life. By 1992 Howard thought enough was known so that he could make experimental contributions toward the prevention of AIDS. HIV and HTLV-1 were known to be lentiviruses, a subtype of retroviruses that encoded several genes in

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Biographical Memoirs: Volume 79 addition to the gag, pol, and env of the simple retroviruses such as SNV. These additional genes were necessary for HIV’s replication. No simple retrovirus had been isolated from people, and no lentiviruses had been isolated from chickens, mice, and cats, which harbor many simple retroviruses. Howard hypothesized that in this current niche of time, human beings have evolved to be resistant to simple retroviruses. A vaccine might be constructed by engineering the gag, pol, and env genes of HIV into a simple retroviral vector and be used to immunize people against infection by HIV (1993,1). Kathy Boris-Lawrie, a postdoctoral fellow, and he began testing this hypothesis by working with the animal lentiviruses, bovine leukemia viruses, and simian immunodeficiency virus, as models. Howard died of cancer in February 1994 before these preliminary experiments were completed. This final project exemplified Howard’s research. It grew out of a thorough appreciation of HIV as a retrovirus. It embodied a bold hypothesis. It will be a lasting sorrow that he did not live to test his hypothesis, to learn from the experiments he proposed, and to contribute his further insights to biology and human welfare. NOTES 1.   G.M.Cooper, R.G.Temin, and B.Sugden, eds. The DNA Provirus: Howard Temin’s Scientific Legacy. ASM Press, 1995. 2.   Dulbecco and Vogt. J. Exp. Med. 99(1954):167–82. 3.   Mannaker and Groupé. Virology 2(1956):838–40. 4.   Bather. Brit. J. Cancer 11 (1957):611–19. 5.   Crawford and Crawford. Virology 13(1961):227–32. 6.   Temin and Rubin. Virology 8(1959):209–22. 7.   Brenner and others. Nature 190(1961):576–81. 8.   Gros and others. Nature 190(1961):581–85. 9.   Goldberg and others. Proc. Natl. Acad. Sci. U. S. A. 48(1963):2094– 2101.

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Biographical Memoirs: Volume 79 10.   Reich and others. Proc. Natl. Acad. Sci. U. S. A. 48(1962):1238–44. 11.   Bader. Virology 22(1964):462–68. Vigier and Goldé. Virology 23(1964):511–19. 12.   Temin. J. Natl. Cancer Inst. 37(1966):167–75. 13.   Kates and McAuslan. Proc. Natl. Acad. Sci. U. S. A. 58(1967):134–41. 14.   Shatkin and Sipe. Proc. Natl. Acad. Sci. U. S. A. 61(1968):1462–69. 15.   Sambrook and others. Proc. Natl. Acad. Sci. U. S. A. 60(1968):1288–95. 16.   Baltimore. Nature 226(1970):1209–11. 17.   Baltimore, Huang, and Stampfer. Proc. Natl. Acad. Sci. U. S. A. 66(1970):572–76. 18.   Temin. J. Natl. Cancer Inst. 37(1966):167–75. 19.   Temin. J. Cell. Physiol. 69(1967):377–84. 20.   Temin. Wistar Inst. Symp. Monogr. 7(1967):103–16. 21.   Temin. Int. J. Cancer 3(1968):491–503. 22.   Figure 11 in Temin. Int. J. Cancer 3(1968):771–87. 23.   Temin. J. Cell. Physiol. 74(1969):9–16. 24.   Temin. J. Cell. Physiol. 75(1970):107–20. 25.   Pierson and Temin. J. Cell. Physiol. 79(1972):319–30. 26.   Smith and Temin. J. Cell. Physiol. 84(1974):181–92. 27.   Temin. Perspect. Biol. Med. 14(1970):11–26. 28.   Temin. Pp. 176–87 in The Biology of Oncogenic Viruses, ed. L.G. Silvestri, vol. 2. North-Holland, 1971. 29.   Huebner and Todaro. Proc. Natl. Acad. Sci. U. S. A. 64(1969):1087–94. 30.   Stehelin and others. Nature 260(1976):170–73; Stehelin and others. J. Mol. Biol. 101 (1976):349–65. 31.   Coffin and Temin. J. Virol. 7(1971):625–34; Coffin and Temin. J. Virol. 8(1971):630–42; Kang and Temin. Proc. Natl. Acad. Sci. U.S. A. 69(1972):1550–54; Kang and Temin. Nature New Biol. 242(1973):206– 208; Mizutani and Temin. J. Virol. 12(1973):440–48; Kang and Temin. J. Virol. 12(1973):1314–24; Mizutani, Kang, and Temin. Cold Spring Harbor Symp. Quant. Biol. 38(1974):289–94. 32.   Temin, Kang, and Mizutani. Pp. 1–13 in Possible Episomes in Eukaryotes, ed. L.Silvestri. North-Holland, 1973. 33.   Temin. Natl. Cancer Inst. Monogr. 52(1979):233–38. 34.   Kelly and Smith. J. Mol. Biol. 51(1970):393–409. 35.   Sharp and others. Biochemistry 12(1973):3055–63. 36.   Kornberg and Baker. DNA Replication. Freeman, 1992. 37.   Southern. J. Mol. Biol. 90(1975):503–17.

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Biographical Memoirs: Volume 79 38.   Maxam and Gilbert. Proc. Natl. Acad. Sci. U. S. A. 74(1977):560–64; Sanger and others. Proc. Natl. Acad. Sci. U. S. A. 74(1977):5463–67. 39.   Hill and Hillova. C. R. Acad. Sci. Paris D 272 (1971):3094–97. 40.   Kang and Temin. J. Virol. 14(1974):1179–88. 41.   Duesberg and Vogt. Proc. Natl. Acad. Sci. U. S. A. 67(1970):1673–77. 42.   Hirt. J. Mol. Biol. 26(1967):365–69. 43.   Kates. Cold Spring Harbor Symp. Quant. Biol. XXXV(1970):743–52. 44.   Keshet and others. Cell 16(1979):51–61. 45.   Shank and others. Cell 15(1978):1383–95 and 1397–1410. 46.   O’Rear and others. Cell 20(1980):423–30. 47.   Williams and Blattner. J. Virol. 29(1979):555–75. 48.   Shimotohno and Temin. Proc. Natl. Acad. Sci. U. S. A. 77(1980):7357–61. 49.   Coffin. J. Gen. Virol. 42(1979):1–26. 50.   Shimotohno and Temin. Figure 2 in Proc. Natl. Acad. Sci. U. S. A. 77(1980):7357–61. 51.   Chen and others. J. Virol. 45(1983):104–13; Wilhelmsen and others. J. Virol. 52(1984):172–82. 52.   Gilmore and Temin. Cell 44(1986):791–800; Gilmore and Temin. J. Virol. 62(1988):703–14; Ballard and others. Cell 63(1990):803–14. 53.   Shimotohno and Temin. Cell 26(1981):67–77. 54.   Watanabe and Temin. Proc. Natl. Acad. Sci. U. S. A. 79(1982):5986–90. 55.   Mann and others. Cell 33(1983):153–59. 56.   Panganiban and Temin. Proc. Natl. Acad. Sci. U. S. A. 81(1984):7885–89. 57.   Drinkwater and Klinedinst. Proc. Natl. Acad. Sci. U. S. A. 83(1986):3402–06. 58.   Coffin. Science 267(1995):483–88. 59.   Wei and others. Nature 373(1995):117–22. 60.   Hu and Temin. Proc. Natl. Acad. Sci. U. S. A. 87(1990):1556–60. 61.   Temin. Proc. Natl. Acad. Sci. U. S. A. 90(1993):6900–6903. 62.   Zhang and Temin. J. Virol. 67(1993):1747–51. 63.   P.Duesberg. Science 241(1988):514. 64.   Temin. Policy Rev. 54(1990):71–72.

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Biographical Memoirs: Volume 79 SELECTED BIBLIOGRAPHY 1958 With H.Rubin. Characteristics of an assay for Rous sarcoma virus and Rous sarcoma cells in tissue culture. Virology 6:669–88. 1959 With H.Rubin. A radiological study of cell-virus interaction in the Rous sarcoma. Virology 7:75–91. 1960 The control of cellular morphology in embryonic cells infected with Rous sarcoma virus in vitro. Virology 10:182–97. 1963 Further evidence for a converted, non-virus-producing state of Rous sarcoma virus-infected cells. Virology 20:235–45. The effects of actinomycin D on growth of Rous sarcoma virus in vitro. Virology 20:577–82. 1964 Homology between RNA from Rous sarcoma virus and DNA from Rous sarcoma virus-infected cells. Proc. Natl. Acad. Sci. U. S. A. 52:323–29. 1967 Studies on carcinogenesis by avian sarcoma viruses. V. Requirement for new DNA synthesis and for cell division. J. Cell. Physiol. 69:53–63. 1970 With S.Mizutani. RNA-dependent DNA polymerase in virions of Rous sarcoma virus. Nature 226:1211–13. With D.Boettiger. Light inactivation of focus formation by chicken embryo fibroblasts infected with avian sarcoma virus in the presence of 5-bromodeoxyuridine. Nature 228:622–24. 1971 The protovirus hypothesis. J. Natl. Cancer Inst. 46:3–7.

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Biographical Memoirs: Volume 79 With J.M.Coffin. Ribonuclease-sensitive deoxyribonucleic acid polymerase activity in uninfected rat cells and rat cells infected with Rous sarcoma virus. J. Virol. 8:630–42. 1973 With N.C.Dulak. A partially purified polypeptide fraction from rat liver cell conditioned medium with multiplication-stimulating activity for embryo fibroblasts. J. Cell. Physiol. 81:153–60. 1974 With G.M.Cooper. Infectious Rous sarcoma and reticuloendotheliosis virus DNAs. J. Virol. 14:1132–41. 1977 With E.Fritsch. Formation and structure of infectious DNA of spleen necrosis virus. J. Virol 21:119–30. 1979 With E.Keshet and J.O’Rear. DNA of noninfectious and infectious integrated spleen necrosis virus (SNV) is colinear with unintegrated SNV DNA and not grossly abnormal. Cell 16:51–61. 1980 With K.Shimotohno and S.Mizutani. Sequence of retrovirus provirus resembles that of bacterial transposable elements. Nature 285:550–54. 1981 With S.K.Weller. Cell killing by avian leukosis viruses. J. Virol 39:713–21. 1982 With K.Shimotohno. Loss of intervening sequences in genomic mouse a-globin DNA inserted in an infectious retrovirus vector. Nature 299:265–68. 1983 With S.Watanabe. Construction of a helper cell line for avian reticuloendotheliosis virus cloning vectors. Mol. Cell. Biol. 3:2241–49.

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Biographical Memoirs: Volume 79 1988 With W.Blattner and R.C.Gallo. HIV causes AIDS. Science 241:515–16. With J.P.Dougherty. Determination of the rate of base-pair substitution and insertion mutations in retrovirus replication. J. Virol. 62:2817–22. With C.Gelinas. The v-rel oncogene encodes a cell-specific transcriptional activator of certain promoters. Oncogene 3:349–55. 1990 With W.-S.Hu. Retrovirus recombination and reverse transcription. Science 250:1227–33. 1993 A proposal for a new approach to a preventive vaccine against human immunodeficiency virus type 1. Proc. Natl. Acad. Sci. U. S. A. 90:4419–20. With J.Zhang. Rate and mechanism of nonhomologous recombination during a single cycle of retroviral replication. Science 259:234–38.

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