Mary Jane Gething on Race for the tPA Clone
  Mary Jane Gething     Biography    
Recorded: 16 Jan 2003

Jim Watson had come back Japan. And there’s this funny story of how, when in Japan Jim was asked to pee in a bowl and he thought—this may be our story, but our imagining was that Jim thought that in collecting Nobel Prize urine, but in fact it turned out that there was a huge effort in Japan to purify to the enzyme urokinase from urine so they were collecting human urine to do it. And Jim’s was just one small part of this.

But anyway Jim thought, you know, he found this out, of course. But he felt, and rightly so, that if Japan was willing to go to that much effort to purify this enzyme there must be something important about urokinase. So he either went especially to talk to Dan Rifkin at NYU or maybe had met Dan Rifkin in the near future after that.

Dan was an expert in these blood clotting and clot breaking enzymes. And Dan told Jim that, yes, but urokinase wasn’t the right enzyme. It didn’t have the specificity of this other enzyme—tissue type plasminogen activator. And that was the one that should be cloned.

And Jim came back and talked to Joe and said, “This is something the lab must be doing because it has billion dollar market,”and—which is has turned out to be, I guess. And so Joe and I started to do this project. In fact, this was even while I was still at Cold Spring…still in London and so it must have been happening when I was coming backwards and forwards, because it was known that this enzyme was a glycoprotein like the one I was working on.

So Joe brought over from Cold Spring Harbor to London a cell line that Dan Rifkin had given us. And some antibody to the protein because Joe had never run a protein gel in his life. So we made cell extracts from these cells and did immunoprecipitations and Ed Harlow helped us at this very early stage. He was a graduate student in London then.

So we started this cloning project with tPA and then at some point, Jim and Joe arranged some funding and, in fact, a company was started up called Cell Biology and then money came from Baxter Travenol through Cell Biology for this project. But it was a very difficult project. Because it was…it turned out to be an extremely stable protein, so we thought because there’s lot of protein there would be lots of RNA but in fact the protein was very stable and there wasn’t much RNA at all. And so it needed a much larger cDNA library to find it in than we expected.

So Doug Hanahan started working with us because—and we would make the, batches of cDNA down in Cold Spring Harbor but it was clear that we needed Doug’s—Doug had even a better liquid gold at that time. So there was this marvelous week when we drove up—Joe and I were going to a Gordon Conference up in New Hampshire.

This would probably be ’82, I should think. So Joe and I had made—I’d made this batch of cDNA which looked better than the ones we’d been doing and, but we had to interrupt to go to this Gordon Conference. I think I was chairing the conference or something; anyway, I had to go.

So we drove up and Doug drove about from Boston and met us in a Howard Johnson’s restaurant on the turnpike and we handed the cDNA over to him and he went back to Boston and spent the next few days transforming it with his absolutely best “Doug Hanahan Five” bacteria or whatever it was, DH-5 alpha and his best liquid gold and we went off to the conference. And then four days later we drove back down—Doug drove out to the turnpike and gave us the transformed library to take down to Cold Spring Harbor.

And then I plated it out, I believe. And then he and I—I remember spending a whole day in the back lab at James plating out multiple copies of this library onto nitrocellulose filters. And that was in the end is a long story which I won’t tell here about Genetics Institute then became involved, a second company, to help with the probing and our library was lost for awhile, I think, while they tried to do their own library, but in the end we jumped up and down and they found it in some deep recess of a freezer. And the clone—the tPA clone, in fact, eventually went quite quickly actually, did come out of our original library that had been backwards and forwards to Howard Johnson’s and so when—I think that was probably a very high point cause that had been a very hard project.

We lost the race, of course. Genentech got the tPA clone some months—two or three months, I believe, before us but it was the clone we went on and did the work later in Dallas to make an improved version of tPA which actually is being sold now. So many years later—it just went to the market last year, I believe.

Mary-Jane Gething, biochemist is Head of the Department of Biochemistry and Molecular Biology at the University of Melbourne where she earned her Ph.D. in Biochemistry in 1974. Subsequently she went to Cambridge to do post-doctoral work.

In 1976, she moved to London to work on protein sequencing and in 1980, Gething and Joseph Sambrook received a NATO grant for travel to collaborate on virus research. She began working at Cold Spring Harbor Laboratory in 1982 where she continued her research of proteins. In 1985, Gething and Sambrook moved to Dallas to work at the University of Texas Southwestern Medical Center. They moved back to Australia in 1994.

Her current research involves protein folding in the cell and the role of molecular chaperone BiP.