1.5 John Coffin — The Origin of Molecular Retrovirology

John Coffin: [00:00:00] I have to start by thanking the organizers, but I would particularly like to thank the Cold Spring Harbor people for the idea of having this meeting. It has been really an enormous honor and pleasure to be able to interact with Bob [Gallo] and with Bruce [Walker] to put it together over the last year or so. I'm going to focus on one very small aspect, but actually very large aspect of [00:00:30] retrovirology in this talk. Before I do that, I want to point out something that—actually to emphasize something that Robin [Weiss] mentioned as well, and that's the importance of Cold Spring Harbor to the field overall.

The annual Retrovirus Meeting, which actually started as the Tumor Virus Meeting and became the RNA Tumor Virus Meeting, and then eventually the Retrovirus Meeting, has been going on [00:01:00] since the late '60s. I've had the honor of being at more of those meetings than any other person, 44 of them altogether, in fact, and every single one since 1975, I had to take a little break as when I was a postdoc in Switzerland and couldn't talk my rather stingy boss into sending me back to the US for this meeting in the meantime.

You can actually judge, in a sense, the progress of the field by scanning [00:01:30] the thickness of the abstract book starting in 1970, going up to the heyday of the oncogenes in here, then the oncogenes left to a separate meeting and the abstract book got quite thin, but surely after that, HIV was discovered and the abstract books rather rapidly get thick again. They've stayed at a pretty good thickness running around 400, 300 to 400 abstracts ever since then.

I first came, as I said, as a graduate student. [00:02:00] To move on more to the topic of the talk: this is [slide] obviously the retrovirus replication cycle that we are all familiar with by the time HIV was discovered. In fact, by the time HTLV (human T-lymphotropic virus) was discovered in 1980, the outlines of this had been pretty much worked out. We could have drawn this same picture and probably did many of them probably did, even before 1980.

But I want you to think for a second [00:02:30] of how the field of AIDS research would have progressed if we had not known this. The answer is it would have been many years. If we started from scratch to figure out retrovirus replication after HIV was discovered as a virus, it would have been many years. The virus would have spread enormously. The toll in human life would have actually been quite substantial if we had not had this information already [00:03:00] at the time HIV was discovered because this information made it possible to take a sample of virus from your culture, which you could see from cytopathic effects with growing virus, do a very simple biochemical assay that any one of you could do overnight and determine that this is almost certainly a retrovirus and in further, pretty much the rest of the replication cycle, at least an outline.

Then that leads to all kinds of other useful information [00:03:30] and eventually, of course, to the rather remarkable treatments that we have for HIV infection today. But what I want to cycle to is really the critical steps in retrovirus replication that characterize retroviruses which are reverse transcription, the formation of the DNA provirus from an RNA template, and the integration step. I will not actually discuss the integration that'll be the next speaker, [00:04:00] Harold [Varmus] will do that. But I do want to talk about reverse transcription and how we came to realize that this is the way retroviruses replicate.

You won't be surprised to hear that this story actually revolves of course, around Howard Temin (1934–1994), who I had the very good fortune by chance. I didn't pick him and he didn't pick me basically, but I [chuckles] ended up in his lab as a graduate student in the late '60s. When I entered the Howard Temin's lab as a graduate student, there were two, sort of [00:04:30] fundamental truths of molecular biology. The first is if it's true for E. coli, it's true for the elephant. That is to say that everything important that you need to know about how genes work, you can learn by studying bacterial systems. This was indeed an article of faith. Eukaryotic systems were just too hard to work with to do the same kinds of elegant experiments you could do with E. coli.

Second, of course, was the Central Dogma [articulated by Crick in 1957]. [00:05:00] That is to say that information flows from DNA. It can be replicated in DNA to RNA, and then certain viruses that can be replicated as RNA to protein, but these arrows are not reversible. This one is obviously not reversible. There actually was no biochemical reason why this one couldn't be reversed, but it was an article of faith among molecular biologists that this is the way it goes. It was enunciated, after all, by Francis Crick (1916–2004, one of the discoverers of the DNA double helix), it had to be true. Even though Crick later denied [00:05:30] that this is what he'd said, but that's an argument for historians to deal with, I think. 

Howard Temin was, as it was pointed out, a graduate student in Renato Dulbecco's (1914–1912, virologist, recipient of 1975 Nobel for work on oncoviruses) lab at Caltech, working with a postdoc there, Harry Rubin (1926–2020, cell biologist and virologist). Harry was a terrific biologist. They developed this, as was pointed out in the previous talk, the focus assay for Rous sarcoma virus. (1) This was the first [00:06:00] quantitative assay for a retrovirus that one could easily do and had very important implications and was led to a lot of the discoveries that Robin [Weiss] mentioned.

But I particularly want to point to one paper. This is in 1960. (2)  Howard was still a graduate student in the lab at the time. He observed, and these are two foci from the paper that he looked at, and he observed that he had different strains. He was using [00:06:30] different strains of the same virus, that differed one from the other genetically somehow. The virus is known at that time to be an RNA virus that had been discovered a few years previously.

He noticed when looking at these foci in the microscope—and you’ll notice that his microscope wasn't as clean as it might've been—that he could see two kinds of foci. Foci of mostly round cells [left on slide; chicken chick fibroblast cells] and foci of mostly elongated spindle-shaped cells [right on slide]. [00:07:00] He called these morph^r for “round,” morph^f for “fusiform.” He discovered that these characteristics breeded true with the virus. If you pick cells from this foci, grew them up, took the virus from them, infected more cells, then you would get cells that looked like this. Even with different cultures of cells and so on. This is not a property of the cells, this characteristic was the property of the viruses.

How, in the discussion of this paper, he wondered, could the genetics of the [00:07:30] virus affect the genetics of the cell? This is an RNA virus. His conclusion was that the present findings indicate that some genetic information responsible for the character of the cell was introduced by the virus. I think at that time he had already intuited exactly what that was, although he was cautious enough that he didn't actually say it. But I'm quite certain that at this point [in 1960?] he had decided for himself that the only way this could be, [00:08:00] was if the virus genetic information was converted from RNA, copied from RNA into DNA, and that DNA must then have been stably associated, i.e, integrated in some way with the cell genome.

The next 10 years were spent trying to elucidate this process and to demonstrate and to prove his ideas. (3) He had some inhibitor studies, which were consistent with the idea that the [00:08:30] virus was being produced from DNA, not from replicating RNA since poliovirus would be was insensitive to Actinomycin D, and also that early infection there is a requirement for DNA synthesis, again, based on the use of inhibitors. Finally, a hybridization study that I'll show you some data from in a second, that led him … to conclude that there actually was DNA [00:09:00] in the cells.

This nature of this data was presented at meetings here [at CSHL]. I have to say that he would come back from those meetings extremely depressed because nobody believed him. His ideas were considered completely heretical because of the power of the central dogma and the difficulty of breaking through the preexisting, ideas of what's going on. The other problem was that none of these data [00:09:30] were all that convincing, quite frankly. In fact, even late in this process of only about half the people in his own lab actually believed in the provirus idea. I forget which side of that poll I was on, actually. For most, actually among the critics of this and the most vocal critics was his old colleague, Harry Rubin.

[00:10:00] The Rubin and Temin feud actually became quite famous among people in the field at the time. This slide actually shows the data from the last paper (4) and showing labeled virus RNA hybridized to infected or uninfected cell DNA. You can see that there seems to be a clear signal in infected cells, and a signal that he ignored actually in [00:10:30] uninfected cells. Other people did not ignore it, they said, "What the hell is going on in these uninfected cells, you must have some background [effect] here, you can't explain, so how do you explain this?" The other problem was he only started with 1,000 counts per minute of a very low specific activity probe. If you do the simple calculation, this amounts to three counts per minute here. This is about 1.2 counts per minute here.

He didn't show [00:11:00] them as counts per 10 minutes or counts for 20 minutes, and Sol Spiegelman (1914–1983, American molecular biologist) was later on to do another famous paper that was alluded to already actually by Robin [Weiss]. What he didn't know and which nobody knew at the time this paper was published was that this was actually a real signal and this was due to endogenous viruses, proviruses in these cells that were only just being discovered at the time this experiment was done.

Enter David Baltimore (b. 1938, co-discoverer of reverse transcriptase with Temin, shared 1975 Nobel with Temin and Dulbecco). David Baltimore and [00:11:30] Howard Temin first met, actually, in 1955, when Howard was a counselor at a summer course at Jackson Labs (Bar Harbor, Maine) and David Baltimore was just entering Swarthmore. Howard had just actually just graduated from Swarthmore as a student. This was, I believe, was their first meeting. David had been studying vesicular stomatitis virus, an RNA virus, and had shown in about 1969, or Alice Huang (Alice S. Huang 黃詩厚, b. 1939, virologist and microbiologist, married David Baltimore in 1968) and his lab had shown about 1969, the vesicular stomatitis [00:12:00] virus can contain an RNA dependent RNA polymerase that was necessary for replication of that virus, [thus] establishing the principle that RNA viruses can contain viral enzymes that are involved in the replication cycle.

Both Howard and David realized that retroviruses could well be doing the same thing, that the enzyme necessary for their replication could in fact, already be present in the viruses, and that led them both to do the same experiment [00:12:30] almost exactly at the same time. And that was to take a preparation of purified virons, in Howard's case, use a detergent to disrupt—my role in this will become clear in a minute—to disrupt the virus and add deoxynucleoside triphosphates (dNTPs), one of which is [radio-]labeled, [plus] magnesium salts, buffer, and so on, and then measure the incorporation of the dNTPs added into DNA. And sure enough you get this nice, robust signal of DNA being made using the viral enzyme and the viral RNA is a template, and you get no signal at all if you don't add virus—or actually heated virus, I think this was here. This experiment [was] again done simultaneously by David Baltimore. Howard first presented at a meeting in May of that year in 1970, at the Tenth International Cancer Congress [in Houston]. (5) Here's his talk here, [00:13:30] “The DNA provirus of Rous sarcoma viruses,” coincidentally chaired by Harry Rubin. If you go back and read Harry's paper, about 50% of that paper [consisted of his] arguments against how the provirus can't possibly be correct.

I very distinctly remember Howard coming back from this meeting, gleefully rubbing his hands, saying how Harry kept making these objections and saying these things that he absolutely knew and demolished with this very simple experiment that he showed here. It was actually [00:14:00] a little bit later was—what really sealed this was the fact that David Baltimore and Howard published exactly the same result at exactly the same time. (6–7) 

This was the Nature [issue] of June 27th, 1970. There's a couple of odd things about this paper, or interesting things about this paper. The first is that if you look at the receipt date, the receipt date here is June 15th, the publication date is June 27, twelve days apart. That's a record actually. [00:14:30] I believe quite certain that's a record for Nature. Watson and Crick I believe was something like 35 days for their paper, Jim [Watson], maybe you need correct me on that. Secondly is Nature took upon themselves to change the order of the authors on this paper. Howard couldn't do anything about that because he actually saw that journal before he saw the proofs.

Again, here, the papers were really almost word for word the same. You'll note that David's paper [00:15:00] did not require the use of the detergent Nonidet or NP-40 and, we always attributed that to the fact that this virus is grown by a government contractor and was beaten up enough that you really didn't need to disrupt the virions to get the enzyme to work [audience laughter]. But this is where my important role in this study comes in, and that is, it was my bottle of NP-40 that was used that Satoshi Mizutani (Postdoc at UW-Madison MacArdle Laboratory for Cancer Research, co-author with Temin on the Nature 1970 article) borrowed to do this experiment. [00:15:30] That's not so trivial! It wasn't that easy to get at the time, you had to write especially and beg for it from Shell Chemical Company, the manufacturer.

This was, in my experience, the most dramatic scientific discovery that I was ever associated with, I'm sure ever will be associated with. It hit the scientific press and the popular press like a bombshell, and I'll come back to why that's probably true. [00:16:00] For over, more than the next year, at least once a month, the front pages of Nature “News and Views” had some sort of breathless headline about the discovery of reverse transcriptase, and all of the studies that were done by a very large number of labs, including a number of people in the audience shortly following that.

It started with this one (8) which actually came from the Houston meeting, it [00:16:30] was somewhat skeptical in fact, and then finally, this breathless one, “Central Dogma Reversed,” (9) and then a whole series of others; “Apres Temin, le Deluge” (10), “Deluge Unabated” (11), “Advice for Teminizers” (12) in which they also use the term “Teminist” in the article itself. This one [“Cancer Viruses: More of the Same”] (13) is actually the first written appearance of “reverse transcriptase” to my mind: it was invented, the term was coined by the at-the-time-anonymous Nature [00:17:00] editorial writer, I'm pretty sure it was John Tooze (b. 1938). In fact, I asked him once it was him and he admitted to it, confessed to it, I should guess I should say, and going on and on. (14–16)

Finally, this one (17), which I already alluded to, is this, the study by Sol Spiegelman and colleagues, including the first author of at least one of these studies. There were several of them was Richard Axel (b. 1946), who became much better known for completely different work later on. [00:17:30] Finally, a year later, we had this, we ended up with this one. (18)

The other important role that I played in this, this also became very well-covered in the popular press. Here's a cover from Newsweek (Feb 22, 1971) that was in about six months afterwards. The other important role that I played in this whole process is if you see this T-flask: I filled that flask with [00:18:00] medium, carefully adjusting the pH so it will properly balance the color of the photograph overall. Actually, I have another cute story regarding a discussion with a photographer but I cannot tell it here. If you press me later on, maybe I will. It was more or less locker room talk actually to be fair.

What did I do after that? Well, one of the most useful things I think I did was this—[00:18:30] this is after I came back actually from my postdoctoral fellowship in the set working independently at Tufts, was to have a long-running collaboration with Bill Haseltine (b. 1944). In fact, I had the longest-running collaboration—the world's longest-running collaboration with Bill, until Bob Gallo and Max Essex came along and stole him away from me. John [M.] Taylor (former postdoc in the Bishop-Varmus lab, now at Fox Chase Cancer Center), and others— it had been observed by a number of people that retroviruses [00:19:00] use a tRNA as a [primer] (19), and I remember at this meeting in talks particularly from John Taylor, although others may have made the same observation, that mapped the binding site for this tRNA down very near the 5’ end of the genome.

How we wondered, could a primer be at the 5' end of the genome, when DNA synthesis proceeds in this direction [from 3' to 5']? We postulated, it had also have been discovered, first by Karin Mölling (b. 1943) I believe, that reverse transcriptase also contains an RNAase H activity, capable [00:19:30] of degrading RNA from DNA-RNA hybrid. And so we postulated that what might happen is that you might have the enzyme starting here at the end of the primer site, and then being able to proceed here by taking advantage of a short redundant sequence in the genome, and that will allow it to essentially jump—although I know some of you hate that term—to jump from one end to the other. [00:20:00] With Bill, who discovered this rather, what he called strong-stop DNA, we showed that the strong-stop DNA was capable of hybridizing tRNA sequences at both ends of the genome, therefore, demonstrating this. That then was the first steps leading to what we understand about the process of DNA synthesis. This was all worked out well before 1980. This was worked out without [00:20:30] molecular cloning, PCR, or facile DNA sequencing more than being able to sequence just a little bit of stuff at a time, and very hard won, I should have to say. (20)

This process then was worked out additionally, but particularly with studies from the Baltimore lab, from the [J. Michael] Bishop (b. 1936) and [Harold E.] Varmus (b. 1939, both awarded 1989 Nobel for discovery of the cellular origin of retroviral oncogenes) lab and a few others, and leading to what we understand for the process of proviral DNA sequences now, and the formation of these long term repeats at the end which provide [00:21:00] an important sync transcriptional control signals for the virus.

As they say, there wasn't much sequencing done. In fact, I did publish a paper, a sequencing paper about this time, the nucleotide sequence at the site of the DNA synthesis, “The 102nd nucleotide is U” (21)—actually, Harold [Varmus] may remember this. This actually has to go on record, although maybe tied with lots of other papers, as the shortest [00:21:30] sequence ever determined, since it's basically a single base, but it was right.

I'll just conclude with a couple of slides. First, why does this have such a dramatic scientific impact? I think there were a couple of reasons, just to philosophize for a couple of minutes. First, they provided dramatic proof of Temin's provirus model. To a one, almost every scientist who had thought about this had sneered at. [00:22:00] Immediately, that got turned around. What was important and part of that was that the experiment had been done simultaneously by two labs at the same time. That made a very big difference I think in the instantaneous acceptance of this. It's also important to know that this experiment was extremely easy to repeat.

In fact, there's a story—and I'm not certain whether it's true or not—but there's a story that when David Baltimore presented the work at the symposium that year here [at CSHL], that [00:22:30] there was one person in the audience, went back to his lab in New York, and came back the next morning to announce to the crowd that he had repeated the experiment. Understanding the replication cycle all of a sudden, provided the key entree into the fundamental basis of cancer and Harold would talk about that in the next talk.

[Reverse transcriptase] quickly also became a very important tool for molecular biology, you may see DNA cloning for example, [00:23:00] as a means a retrovirus assay and discovery, and this was, as we've already heard something of a two-edged sword. Quite a few rumored viruses of the sort were in fact discovered this way because the difference in these assays between reverse transcriptase, et cetera, polymerase, and so on was not well clarified.

In fact, as I said this discovery of HTLV-1, the discovery of HIV were really only made possible by having these [00:23:30] reverse transcriptase assay ready to go, so that you could do it in a couple of hours and determine that your virus was indeed a retrovirus. Finally, as a target for any antiviral drugs, we all know that current anti-HIV therapy rests primarily on the use of reverse transcriptase inhibitors, not solely but primarily. A triplet is the single-pill regimen is all reverse transcriptase inhibitors. [00:24:00]

However, in the early days, there was a lot of work done, in completely misguided experiment on the idea of using reverse transcriptase inhibitors in potentially virus-induced cancers, which actually made no sense whatsoever. I never did understand why the government was pouring so much money into it for that reason. I think it did provide important basis for later on the discovery about use of these [00:24:30] compounds as anti-HIV agents.

Finally, why all the hype, the non-scientific hype? Well, for one thing, it injected new life into the cash-rich but idea poor Special Virus Cancer Program at the time. Although, you could argue that it did not necessarily lead to a very scientifically efficient use of money. I think it provided an important impetus not the sole impetus for Nixon's war on cancer [00:25:00] in 1971. Mary Lasker was probably the sole primary impetus.

It was touted as providing a possible path to cancer vaccine and cure, which of course, never really came to be because the virus that then Bob Gallo discovered in 1980, the first oncogenic human retrovirus, also turned out to be the last oncogenic human retrovirus. There just aren't that many of them.

The Zeitgeist on college campuses. [00:25:30] 1970 was a year of tremendous ferment. The president's office at MIT was occupied. There was a bombing at the University of Wisconsin, killing a postdoc in the physics building. Three students were killed in Kent State.  There was tear gas up and down the streets of university campuses. It was a very dramatic time in that respect. Finally, the sheer drama of a lonely scientist with a brilliant but heretical insight, finally coming up with a simple proof of his ideas after a decade of [00:26:00] struggles against the wall of skepticism, I think was a very important part of the overall historical picture that we need to keep in mind here.

Harold [Varmus], the next speaker, will speak about integration. I just showed you one thing to introduce that. This is my slide of mechanism of integration I drew about 1985 showing-- and this was all understood. The structure of the integrated provirus was known, but almost nothing was known about the mechanism, and that's why I put in this, "a [00:26:30] miracle occurs" slide. Finally, in a tragic note at the end, as I'm sure you're all aware, Howard Temin died February 9, 1984. You may not be aware—many of you knew Sheldon Wolff (1930–1994) was a very important figure in NIAID, he was the clinical director before he became Chair of Medicine at Tufts. He had some very important roles in getting the AIDS research program going in its early days. He died on [00:27:00] exactly the same day, just coincidentally. Thank you.

[applause]

Thank you. You should get a microphone. 

Julie Overbaugh: Your talk was so nicely about the story of Temin and being the person who just went doggedly against this set of the [00:27:30] field. The way we do science now is almost the opposite. We try to have consensus and we have big programs. Just thinking back and looking then and now do you think we're missing opportunities by not having that kind of science?

John Coffin: I think that's entirely possible. I think there's still room for heresy in science. I think there's a need for heresy in science to be honest with you. You have to pick your heretic though. A lot of guys who are talking heresy are just screwballs. [00:28:00] [chuckles] It can be hard for you and me to decide that sometimes. There absolutely is, and there needs to be a room in scientific funding still for really basic, oddball-sometimes ideas that seems to be getting lost now in the way scientific review is done, the way funding decisions are made and [00:28:30] the large team science programs which the funding institutes like, but it may not be the best way for—there's certainly a need for that kind of thing to go on, but there's still a need to keep funding only individual scientists with screwball ideas.

Steve Goff (moderator): I remember one interesting thing that reminds me of-- a story that reminded me of which was when RNA splicing was discovered.

John: Also, first presented here [00:29:00] at a Retrovirus Meeting.

Steve: All true. The very next day, almost 10 people came out of the woodwork, pulling films out of their drawers that they showed, "Well, I knew about splicing too", but they never had figured out what this film was meant until it was made apparent. [chuckles]

John: I had a few films like that that I never understood until later on either, although it wasn't splicing. We had done experiment more; we could have discovered splicing but [00:29:30].

Bob Gallo: Another good example, maybe, but certainly not in the same category as Howard, but Ludwik Gross (1904–1999) in the 1950s with the first mammalian leukemia virus. If we remember the chairman at the University of Chicago and Medicine told me that when he would go to the Federation meetings in those days, everybody used to leave the road that he set down in, because he was working in the Bronx VA, originally out of the trunk of his car with his mice. He was making these claims that the mouse get leukemia from the mother and it was something transmissible and it was the virus-- [00:30:00] 

John: This meeting, the  tumor virus meeting used to be DNA tumor viruses and then RNA tumor viruses. The first speaker in the RNA tumor virus part of it usually couldn't be heard from the sound of the automobiles starting up and leaving from all those people from the first half of the meeting.

[00:30:19] Ruth Ruprecht: John, this was a really enjoyable summary of the history. I want to congratulate you on one of the highlights: that bottle of NP-40.

[laughter]

I actually joined that mysterious lab in New York, Sol Spiegelman's [lab], a few weeks after that telephone call that you told about. So the story was still fresh. Spiegelman's people at the time worked with two different viruses, looking at that same incorporation of DNA precursors. They worked with AMV [avian myeloblastosis virus] and with RoV [rotavirus]. And they had no mysterious bottle of NP-40. So with AMV, nothing got in: the virus was too dense, too intact. With RoV, everything was chewed up, so no incorporation. When he heard about your wonderful bottle of NP-40, all they had to do back in New York was add some of that mysterious thing and viòla.

[00:31:19] John: Lot's of detergents work, of course, as it turns out.

Steve: Okay, any more? All right, I think we should, probably in the interest of time, maybe stop. One more, Jonathan gets a call.

[00:31:33] Jonathan Stoye: I've got to ask John a question. You said that if we hadn’t had the reverse transcriptase of the life cycle worked out in 1982 or 1983, it would have been much longer until we understand how retroviruses work. If we'd had everything that came for the molecular biology, if we'd started in 1983, wouldn't we have figured out, very quickly, how the virus replicated?

[00:31:59] John: I think if we knew then, everything else that we knew about molecular biology except for the retrovirus replication cycle,—I haven't done the detailed research on just exactly what we knew and we didn't—I would postulate that it would have been a much longer time before we were able to deal with HIV. Bob, would you disagree with that?

[00:32:30] Bob Gallo: Absolutely agree with you. I agree with you very much.

[00:32:43] [END OF AUDIO]

Citations

1. Temin, Howard M., and Harry Rubin. “Characteristics of an Assay for Rous Sarcoma Virus and Rous Sarcoma Cells in Tissue Culture.” Virology 6, no. 3 (December 1958): 669–88. doi:10.1016/0042-6822(58)90114-4.

2. Temin, Howard M. “The Control of Cellular Morphology in Embryonic Cells Infected with Rous Sarcoma Virus in Vitro.” Virology 10, no. 2 (February 1960): 182–97. doi:10.1016/0042-6822(60)90038-6.

3. On Temin's attempts to demonstrate reverse transcription:

• Temin, Howard M. “Homology Between RNA from Rous Sarcoma Virus and DNA from Rous Sarcoma Virus-Infected Cells.” Proceedings of the National Academy of Sciences 52, no. 2 (August 1, 1964): 323–29. doi:10.1073/pnas.52.2.323.
• ———. “The Effects of Actinomycin D on Growth of Rous Sarcoma Virus in Vitro.” Virology 20, no. 4 (August 1963): 577–82. doi:10.1016/0042-6822(63)90282-4.
• ———. “The Participation of DNA in Rous Sarcoma Virus Production.” Virology 23, no. 4 (August 1964): 486–94. doi:10.1016/0042-6822(64)90232-6.
  

4. Temin, Howard M. “Homology Between RNA from Rous Sarcoma Virus and DNA from Rous Sarcoma Virus-Infected Cells.” Proceedings of the National Academy of Sciences 52, no. 2 (August 1, 1964): 323–29. doi:10.1073/pnas.52.2.323.

5. For the conference proceedings see: Clark, Randolph Lee, ed. Oncology, 1970: Being the Proceedings of the Tenth International Cancer Congress. Chicago: Year Book Medical Publishers, 1971.

6. Nature 226, June 27, 1970:

• Baltimore, David. “Viral RNA-Dependent DNA Polymerase: RNA-Dependent DNA Polymerase in Virions of RNA Tumour Viruses.” Nature 226, no. 5252 (June 27, 1970): 1209–11. doi:10.1038/2261209a0.
• Mizutani, Satoshi, and Howard M. Temin. “Viral RNA-Dependent DNA Polymerase: RNA-Dependent DNA Polymerase in Virions of Rous Sarcoma Virus.” Nature 226, no. 5252 (June 27, 1970): 1211–13. doi:10.1038/2261211a0.
  

7. For an alternate account that suggests Baltimore and Temin worked more independently, see Judson, Horace Freeland. “No Nobel Prize for Whining.” The New York Times, October 20, 2003, sec. A, page 17, https://www.nytimes.com/2003/10/20/opinion/no-nobel-prize-for-whining.html

8. Our Cell Biology Correspondent (likely John Tooze, see above). “Cancer: DNA from RNA Template.” Nature 226, no. 5250 (June 13, 1970): 1003–4. doi:10.1038/2261003b0.

9. “Central Dogma Reversed.” Nature 226, no. 5252 (June 27, 1970): 1198–99. doi:10.1038/2261198a0.

10. “Après Temin, Le Déluge.” Nature 227, no. 5262 (September 5, 1970): 998. doi:10.1038/227998a0.

11. “Deluge Unabated.” Nature 228, no. 5270 (October 31, 1970): 410–410. doi:10.1038/228410a0.

12. “Advice for Teminizers.” Nature 227, no. 5261 (August 29, 1970): 887–887. doi:10.1038/227887a0.

13. A Special Correspondent (John Tooze). “Cancer Viruses: More of the Same.” Nature 227, no. 5261 (August 29, 1970): 887–88. doi:10.1038/227887b0.

14. Our Cell Biology Correspondent (John Tooze). “Reverse Transcriptase: Transforms Initiated.” Nature 230, no. 5295 (April 23, 1971): 493–94. doi:10.1038/230493b0.

15. Our Cell Biology Correspondent (John Tooze). “Reverse Transcriptases: Roundabouts and Swings.” Nature 228, no. 5278 (December 26, 1970): 1255–1255. doi:10.1038/2281255a0.

16. “Calling the False Reverse Transcriptases.” Nature 231, no. 5301 (June 4, 1971): 283–283. doi:10.1038/231283b0.

17. “Reverse Transcriptase in Human Milk Virus.” Nature 231, no. 5298 (May 14, 1971): 80–80. doi:10.1038/231080a0.

18. “Happy Birthday, Reverse Transcriptase?” Nature New Biology 231, no. 23 (June 9, 1971): 161–161. doi:10.1038/newbio231161a0.

19. Note: this is a correction from John Coffin (email to Mila Pollock July 9, 2020).

20. Coffin, John M., and William A. Haseltine. “Terminal Redundancy and the Origin of Replication of Rous Sarcoma Virus RNA.” Proceedings of the National Academy of Sciences 74, no. 5 (May 1, 1977): 1908–12. doi:10.1073/pnas.74.5.1908.

21. Coffin, John M., and William A. Haseltine. “Nucleotide Sequence of Rous Sarcoma Virus RNA at the Initiation Site of DNA Synthesis: The 102nd Nucleotide Is U.” Journal of Molecular Biology 117, no. 3 (December 15, 1977): 805–14. doi:10.1016/0022-2836(77)90071-7.

Index

• 1.1 James D. Watson — Welcome
• 1.4 Robin Weiss — Retrovirus History and Early Searches for Human Retroviruses
• 1.6 Harold Varmus — Animal Retroviruses and Cancer Research
• 1.7 Max Essex — From Feline Leukemia Virus to AIDS in Africa
• 2.4 Robert Gallo — Discoveries of Human Retrovirus, Their Linkage to Disease as Causative Agents & Preparation for the Future
• 4.0.2 Ruth Ruprecht — Session 4, Introduction 2
• 6.3 Bruce Walker — Role of T Cells in Controlling HIV Infection
• 8.6 David Baltimore — Bringing it to an End (And Where Are We Going?)
• 10th International Cancer Congress, Houston, October 1970
• activism, civil rights, protests, and social movements
• Annual meeting on retroviruses, CSHL
• avian myeloblastosis virus (AMV)
• Axel, Richard (b. 1946)
• Bishop, J. Michael (b. 1936)
• Caltech (California Institute of Technology)
• cell culture, tissue culture, immortalized cell line
• Central Dogma
• chicken
• Cold Spring Harbor Laboratory (CSHL)
• counterfactual history
• diagram
• Dulbecco, Renato (1914–2012)
• E. coli
• endogenous retrovirus (ERV)
• epistemic object becomes the technical object
• focus assay for Rous sarcoma virus
• funding and grants
• gene mapping
• graduate training and early career in science
• Gross, Ludwik (1904–1999)
• Haseltine, William A. (b. 1944)
• HTLV (human T-lymphotropic virus)
• iconoclasm in science
• Jackson Laboratory
• Kent State massacre, May 4, 1970
• leukemia and lymphoma
• LTR (long terminal repeat)
• memory
• microscope — electron and optical
• MIT (Massachusetts Institute of Technology)
• Mizutani, Satoshi
• models (model systems, model organisms, modeling)
• molecular cloning
• Mölling, Karin (b. 1943)
• National Institute of Allergy and Infectious Diseases (NIAID)
• Nature (journal)
• New York
• Newsweek
• non-nucleoside reverse-transcriptase inhibitors (NNRTIs)
• Nonidet P-40 (NP-40)
• nucleic acid hybridization
• nucleosides, nucleotides, nucleoside analogues, nucleoside reverse transcriptase inhibitors (NRTIs), nucs
• observation
• oncogene (onc)
• PCR (polymerase chain reaction)
• politics of scientific journal publishing
• Pollock, Ludmilla "Mila"
• provirus
• radionuclide, radiolabeling, radioactive tracer
• reproducibility; experimental reproduction
• reverse transcriptase
• reverse transcriptase assay
• ribonuclease (RNase); RNase H
• RNA polymerase, RNA replicase
• rotavirus
• Rous sarcoma virus (RSV)
• Rubin, Harry (1926–2020)
• scientific controversy and consensus
• sequencing
• Session 1: The Story of Animal Retroviruses
• Shell Chemical Company
• simultaneous discovery (multiple discovery)
• Special Virus Cancer Program (SVCP), 1964–1978
• Spiegelman, Sol (1914–1983)
• statistical measurement
• Sterling Hall bombing, August 24, 1970
• Stoye, Jonathan P. (b. 1952)
• Swarthmore College
• Switzerland
• Taylor, John M.
• Temin, Howard M. (1934–1994)
• Tooze, John (b.1938)
• University of Wisconsin-Madison
• vesicular stomatitis virus (VSV)
• War on Cancer, 1971– 
• Wolff, Sheldon M. (1930–1994)