Salary History from Grad School in 1966 to 2010 – No, I didn’t get rich.
The Garret Ihler Story – My Grad School Mentor
Who was the first to isolate a gene?
It was Garret Ihler in Charlie Thomas’ lab at Harvard in 1968-69 before the advent of recombinant DNA. The paper, appropriately titled “Isolation of Pure lac Operon DNA”, was published in Nature (vol. 224 pages 768-774) in 1969. This paper, certainly, was fresh in the minds of Dan Nathans and Hamilton Smith a few years later as they developed restriction enzymes to cut and paste defined DNA fragments (Nobel Prize for Medicine in 1978). Ihler’s work started molecular biologists thinking of the benefits of molecular cloning of recombinant genes. Although lac is an E. coli (lactose) operon consisting of three structural genes (encoding the enzymes required for metabolism of lactose), lac is a single transcriptional unit with one promoter and two regulatory domains (for the repressor and operator). Ihler’s paper points out that purification of individual genes would permit investigation of their mechanisms of transcriptional control and expression (the subject of my Ph.D. Thesis). To put this event in perspective, the hottest area of genetics at that time was transcriptional control in the E. coli bacterium with the discovery of sigma factor and investigations into the mechanism by which RNA polymerase transcribed genes. A few years later we would be able to cut genes with restriction enzymes and paste these genes with a ligase into vectors that could be used to transfer genes into cells. Nathans and Smith’s work allowed Paul Berg to put the E. coli gene gpt (encoding guanine phosphoribosyltransferase) next to the SV40 promoter to correct the genetic defect for Lesh-Nyan Syndrome (Nobel Prize in Chemistry, 1980). I benefited from all four of these developments because through their work I had the incentive and was able to clone the human beta-actin gene in December of 1982 and characterize its function in recipient cells using Paul Berg’s vector. Garret Ihler’s work and his mentoring instilled in my mind the goal and benefits of purifying a gene. There is more to the story of Garret Ihler’s accomplishment.
If you look at the 1969 Nature paper, you will find that it has 6 authors – Shapiro, MacHattie, Eron, Ihler, Ippen, and Beckwith. If you look at the acknowledgement at the end of the paper, you will see that Garret Ihler was actually a postdoctoral fellow in Charlie Thomas’ lab and that Thomas’ grant supported the work. In fact, the lac operon was isolated and purified in Charlie Thomas’ lab (noted for DNA replication research) by Garret Ihler.
Picture the group in Beckwith’s lab pondering the prospect of isolating a pure gene but not knowing how. Shortly afterwards Karin Ippen takes a stroll across campus with Ihler describing to him the issue of the day in the Beckwith lab. Picture Ihler enamored with Karin Ippen (the feeling turned out to be mutual). He was also clever especially with lambda transducing phages – these phages with the transduced lac operon from E. coli had been isolated in the mid-1960’s. In fact Ihler could take these phage strains “off the shelf” at a moment’s notice with the lac operon inserted into the lambda genome in opposite directions. Shortly Ihler had thought of the solution for the Beckwith lab with Ippen as his audience. He had the tools at his fingertips – the strains of transduced lac, a reliable exonuclease, and a method of separating and purifying each DNA strand from the lambda double helix. Nevertheless, he offered the idea to Ippen who took it back to the Beckwith lab; subsequently, she was pushed aside by the aggressive members of the group. So Ihler decided to act quickly. He grew up two transducing phages each with the lac operon in opposite orientations, he separated the DNA strands using poly-UG, and then hybridized the two opposite strands. He isolated the “heavy” DNA lambda phage strands (higher poly-UG binding strands) from two different phages each with the opposite lac orientation. When preparations of the two heavy strands were allowed to hybridize only the lac operon sequences could hybridize – the two strands were complimentary only in this part of the phage genome because the flanking lambda DNA sequences of the hybrid DNA were from the same heavy strand of the lambda genome and thus homologous, not complimentary sequences. Ihler nibbled away the lambda DNA “tails” with an exonuclease and precipitated the remaining DNA to yield a pure DNA fragment encoding the lac operon.
Today this seems quite a simple experiment (if you are a molecular biologist), but in the context of the late 1960’s it was a clever experiment with a conceptual pay-off. What happened next poisoned the environment for molecular geneticists especially in the Boston area.
Beckwith took this work and euphorically ran with it. Four co-authors from the Beckwith lab were added with trivial contributions and the paper was published. Beckwith scheduled a news conference basically to take credit for the work much to the embarrassment of Ihler. He used this news conference to belittle the accomplishment and used this media platform to express his personal political agenda which had no connection to Ihler’s work. Surprisingly, the tone of this press conference is preserved in a paper by Beckwith published a year later (Bacteriological Reviews 34:222-227, 1970) where he underplays the role of Ihler – he contributed “a critical idea”. When I read the final section of this paper I could only conclude that Beckwith was deluded by his perception of self-importance. This paper can be found on the Internet for anyone wishing to read it along with Ihler’s recounting of the story at http://mbch.tamu.edu/ihler/ .
Luckily for me, Ihler took a faculty position at the University of Pittsburgh, School of Medicine, and earned his M.D. at the same time. He and Karin married while I was finishing up my Ph.D. He later became an esteemed Professor at Texas A&M noted and especially appreciated by medical students for his stimulating lectures. Karin also developed a productive research program as a Full Professor of Microbiology and Genetics at A&M. Sadly Karin died of breast cancer in 1995 with Garret at her side. Her presence and work has been memorialized through the annual Karin Ippen-Ihler Lecture Series at A&M.
Ihler was probably correct in his belief that Beckwith’s press conference led to misplaced fears and “widespread attempts to regulate cloning and gene transfer”. It did not help that Michael Crichton published “Andromeda Strain” in the same year. Fortunately, all of this concern had fallen by the wayside 13 years later when I was doing this type of work. But now we are experiencing the same hysteria over stem cell research.
Some Trials and Tribulations at Johns Hopkins
I was reminded of the following experiences during my six and a half years at Johns Hopkins by the news in 2006 of fraud by the Korean stem cell scientists who made worldwide news in May of 2005 for their published cloning of human stem cell lines corresponding to afflicted human individuals. Although it seems certain that the Koreans’ work will be disproved and retracted, I prefer to wait for the dust to settle before commenting further on this fiasco. However, in this context I thought that I could discuss my own brushes with overzealous ambition and fraud that occurred during my 30 year career as a bench scientist and researcher. Here is part 1 which relates to events of the 1970’s at Johns Hopkins. In part 2, I follow-up with some more bizarre examples that occurred in the 1980’s while I was at NIH and then at the Pauling Institute. Read the rest of this entry »
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