Recorded: 01 Jun 2001
Harry Noller: …I would say I’m more like Winship—you know, intuitive. You get these methods working and you want to try them out on something and of course you want to try them out on something interesting because when you get through, it would be a cool answer that you’ll be able to enjoy as well as just the, you know… So I was as much interested in the technology as in the biology. I had it on both sides; in the MRC, there were [Sydney] Brenner and [Francis] Crick who are totally into the biology, the methods were there in the service of the biology, whereas Hartley and [Fred] Sanger were totally into the methods. Fred Sanger, who is a hero of mine and Winship’s. This is a guy who worked every day in the lab his whole life. Even after he had two Nobel Prizes, he was coming to the lab and working all day and you’d have to stand there and talk to him at the lab bench if you wanted to talk to him. But he was absolutely only interested in the methodology.
Winship Herr: Well, he was interested in the methodology and moving—and pushing the envelope.
Harry Noller: Yeah, of course, in the most creative way you can imagine.
Winship Herr: And then, he enjoyed sort of overlapping genes or new genetic codes or….
Harry Noller: These were a little chuckle for Fred. He was not beating his chest at all when he found overlapping genes in phi x and this was kind of: “That will amuse the chaps on the second floor.”
Winship Herr: So, if I can follow-up now—you went, you did go—you saw the ribosome as protein chemistry problem, that you could provide something with your expertise….
Harry Noller: That I’d be able to help, yeah.
Winship Herr: And clearly that switched with time—and you ended up focusing on the RNA. But you knew going in it had RNA and proteins. And in fact in the late—originally when Jim [Watson] was excited about the ribosome—pre-messenger RNA—the excitement was that maybe the RNA was the coding RNA in the ribosome. And then when it wasn’t the coding, it became, all of a sudden, less interesting because people didn’t think of the RNA as the enzymatic function.
Harry Noller: It was essentially viewed as a carrier for ethanol precipitations.
Winship Herr: So when—there’s the kethoxal experiment where you inactivated the ribosome and it was the RNA that had been inactivated and not the protein. But even in the introductory paragraph—which I didn’t show in the introduction—I mean, you speak about how there are lots of studies on the ribosomal proteins and conspicuously few on the RNA, even though it’s two-thirds of the mass and it’s probably going to do something. But do you remember, sort of, where along the way you had an appreciation that the RNA was being ignored, or did it—were you forced into it by the kethoxal experiment?
Harry Noller: It was absolutely out of our data. Because I was so convinced that we were going to knock out the function by modifying the proteins. I absolutely bet the farm on this. It was my whole game plan for my, the first years of my career if not longer, and I was frustrated because we were throwing everything but the kitchen sink at the ribosome—at the proteins—and nothing was happening until Rose Bengal, like you said. And then when we did the reconstitutions, the RNA was hit but I didn't believe it was—I thought it was structural damage that it was causing. I didn't think the proteins, I didn’t think the RNA was really being functionally hit, but it was just, sort of out of, you know that when you talk about these things: “Well, you gotta do the control experiment.” Let’s take a reagent that should directly hit the RNA since Rose Bengal is known to hit guanine, we later found in the literature. Then, let’s try to find a guanine-specific reagent; here is kethoxal, which was perfect, it was very specific model conditions, and seemed to be quite specific for guanine.
And then here was this kid, Brad Chaires, who had been screwing around his whole senior year with Hogan, and needed a senior thesis bad, in a hurry. And I said, “Well, here. Why don’t you try this reagent?” and he threw it in with ribosomes and that day he showed me, I mean, they were inactive. And I went “Whoa.”
Winship Herr: That inactivation curve was very fast, right? Rose Bengal also was that fast?
Harry Noller: Yeah, sort of. Yeah, on that rough time scale, a halftime of at fifteen minutes or so. But I didn’t believe it at first. Cause, here’s a guy who hardly has been in a lab before and he’s taking ribosomes and they’re inactive. Well, there’s a lot of ways to inactivate the ribosome. And the curve was all—it didn’t look like the one you saw [indicates up and down] and so he went in and did it again and again, and the curves got better and became more convincing…and I did it and got it and we started believing it and then we did the protection experiment with tRNA and tRNA protected. And then I said, “Well, are we hitting like a hundred guanines, are we just destroying the ribosome?” and then we saw—when we did it with radioactive kethoxal, that we were only hitting about ten or something like that out of five hundred, or four hundred guanines, and you could protect by protecting about a half dozen. So, so the conclusion was we were hitting about six guanines and killing the ribosome. So that was—that made me a believer. And then there were two experiments that came out, actually, from Nomura’s lab and from Jim Dahlberg’s lab that made me, that really supported this. One was actually spectacular. It was the Colicin E3 and that was published around the same time.
Winship Herr: You reference it.
Harry Noller: Yeah, I think the year was 1971. So Colicin is this protein that is an endonuclease and it cuts one bond in the RNA that between, as it turns out, 1493 and 1494. We know right exactly what they’re doing now and it kills the organism by killing the ribosome. So you cut one bond in a 16S RNA and the ribosome is dead. Okay? And the other thing was the resistance to kasugamycin, which was due to absence of methylation of two adenines near the 3’-end of the RNA in that loop. And so that was published, I think, in 1972.
Winship Herr: I think you reference it also.
Harry Noller: So when you read those papers they are interpreted completely in terms of the protein paradigm. In the Colicin paper they say, “Oh I think we’re losing protein P-15, which binds near the end, so that’s why it’s down.” It was not even. “Maybe it’s the RNA.” It wasn’t even discussed, which shows you how deep the protein paradigm was.
Winship Herr: But it’s interesting to me, because I actually took your biochemistry course, which was the fall of my junior year, which was ‘72 when McGovern was running for president. You gave us the day off on Election Day to go campaign or whatever. And that fall a lot of your lecture was on reconstitution, talking about Nomura’s reconstitution pathway, and all the proteins. It was still very protein-dominated, but of course, that was just when you were, at the front lines, you were doing the inactivation of the RNA.
Winship Herr, director of the University of Lausanne School of Biology and member of EMBO. He earned his Bachelor of Arts degree at the University of California in 1974 and Ph.D. for studies on recombinant retroviruses in leukemogenic mice with Walter Gilbert from Harvard University in 1982. He completed his postdoctoral research studies in Cambridge (England) with Frederick Sanger and with Joe Sambrook in Cold Spring Harbor. After that he joined the Cold Spring Harbor Laboratory faculty in 1984. From 1994 till 2002 he was an assistant director of the Laboratory and founding dean of the Watson School of Biological Sciences from 1998 till 2004. He is a professor of the Center for Integrative Genomics at the University of Lausanne.
Winship Herr is a former National Science Foundation predoctoral fellow, Rita Allen Foundation Scholar, Helen Hay Whitney postdoctoral fellow, and Lita Annenberg Hazen Professor of Biological Sciences.
Harry Noller, is best known for his work on on ribosomal RNA structure and function, currently the director of the University of California, Santa Cruz's Center for the Molecular Biology of RNA. He received his B.S. in biochemistry at the University of California, Berkeley and his Ph.D. in chemistry from the University of Oregon.
He received the Rosenstiel Award for Distinguished Work in Basic Medical Sciences together with Drs. Moore and Steitz for their research on the ribosome. Harry Noller has been awarded Paul Ehrlich and Ludwig Darmstaedter Prize granted by the Paul Ehrlich Foundation.
He is a member of National Academy of Science, RNA Society and American Academy of Art and Science.