At that dicey moment in history, Sogin found himself, by age and geography, at the top of the rolls for his local Selective Service board. He hadn’t been drafted yet, but it seemed imminent, and this was before the first lottery made draft boards less arbitrary. “I had to make a sudden decision whether to stay in school or whether to go to Vietnam.” The war was at its ugliest; the Tet offensive in February that year had curdled the thinking of many young American males (including Mitch Sogin and me), and, unfair as it was, you could still get a deferment for graduate school. “Decided to stay in school,” Sogin told me. “It was simple.” He began work toward a doctorate under the mentoring of Woese. His topic was ribosomal RNA.
Woese had noticed something about Mitch Sogin during their early interactions: the kid was not just smart but also handy around equipment. Some combination of talents—dexterity, mechanical aptitude, precision, patience, a bit of the plumber, a bit of the electrician—made him good not just at experimental work but also at creating the tools for such work. Sol Spiegelman had ordered and paid for a collection of apparatus to be used for RNA sequencing by the Sanger method; but now Spiegelman was off to Columbia, leaving behind the tools.
“So Carl inherited that equipment. But he had no one that knew how to use it.” No one, that is, until Sogin joined his lab. “I was essentially responsible for importing all the technology”—importing it from Spiegelman’s lab, and other sources, into the Woese operation. Sogin learned as much as possible from Bishop about Fred Sanger’s techniques before Bishop decamped to New York, and then Sogin became Woese’s handyman as well as his doctoral student, assembling and maintaining an array of hardware to enable the sequencing of ribosomal RNA.
Woese himself was not an experimentalist. He was a theorist, a thinker, like Francis Crick. “He never used any of the equipment in his own lab,” Sogin said. None of it—unless you count the light boxes for reading films. Sogin himself had built these fluorescent light boxes, on which the film images of RNA fragments, cast by radioactive phosphorus onto large X-ray negatives, could be examined. He had converted an entire wall of bookshelves, using translucent plastic sheeting and more fluorescent bulbs, into a single big, vertical light box, like a bulletin board. They called it the light board. Viewed over a box or taped up on the light board, every new film would show a pattern of dark ovals, like a herd of giant amoebae racing across a bright plain. This was the fingerprint of an RNA molecule. Recollections from his lab members at the time, as well as a few old photographs, portray Carl Woese gazing intently at those fingerprints, hour upon hour.
“It was routine work, boring, but demanding full concentration,” Woese himself recalled later. Each spot represented a small string of bases, usually at least three letters but no more than about twenty. Each film, each fingerprint, represented ribosomal RNA from a different creature. The sum of the patterns, taking form in Carl Woese’s brain, represented a new draft of the tree of life.
The mechanics of this effort in Woese’s lab, during Mitch Sogin’s time and for much of the next decade, were intricate, laborious, and a little spooky. They involved explosive liquids, high voltages, radioactive phosphorus, at least one form of pathogenic bacteria, and a loosely improvised set of safety procedures. Every boy’s dream. Courageous young grad students, postdocs, and technical assistants, under a driven leader, were pushing their science toward points where no one, not even Fred Sanger or Linus Pauling, had gone before. The US Occupational Safety and Health Administration (OSHA), though recently founded, was none the wiser.
The fundamental goal was to sequence variants of a molecule from the deepest core of all cellular life, compare those variants, and deduce the history of evolutionary relationships since the beginning. Woese had already settled on that one universal element of cellular anatomy, the ribosome, the machine that turns genetic information into proteins, but there remained a crucial decision: Which ribosomal molecule should he study? Ribosomes comprise two subunits, as I’ve mentioned—a small one snuggled beside a larger one, like an auricle and a ventricle of the heart, each constructed of both RNA and proteins. The RNA fractions include several distinct molecules of different lengths. At first, Woese targeted a short RNA molecule from the large subunit, known as 5S (“five-S”) for obscure reasons that I don’t ask you to contemplate. Just remember 5, a smallish number. That molecule proved unsatisfactory because its very shortness limited the amount of information it contained. The alphabet of nucleotides composing RNA is slightly different from that of DNA—it’s A, C, G, and U (for uracil) in place of T (for thymine)—and there was just not enough of the A-C-G-U alphabet in any little 5S sequence to distinguish different creatures from one another. So he switched to a longer molecule in the small subunit, and at the risk of causing your eyes to roll back in your head, I’m going to tell you its name. Why? Because it’s important, and once you’ve got it, you own it: 16S rRNA. There. Not so bad?
In English we say: “sixteen-S ribosomal RNA.” It’s a structural component of every bacterium on Earth, and bacteria were what Woese studied initially.
There’s a close variant, 18S rRNA, in the ribosomes of more complex creatures, such as animals and plants and fungi. This 16S molecule and its 18S variant, therefore, could serve as the reference standard, the great clue, for deducing divergence and relatedness among all cellular organisms. It was, arguably, the single most reliable piece of evidence, molecular or otherwise, for drawing a tree of life. And that recognition, though it never made the front page of the New York Times, was Carl Woese’s single greatest contribution to biology in the twentieth and twenty-first centuries.
The immediate goal for Woese, back in the early 1970s, was to extract ribosomal RNA from different organisms, to learn as much as possible about the genetic sequence of the chosen rRNA molecule from each organism, and to make comparisons from which he could gauge degrees of relatedness. He started with bacteria, because many kinds of bacteria are easy to grow in a lab, and their collective history is very ancient. Looking at bacteria from numerous different families allowed him the prospect of seeing contrasts, even in such a slowly evolving molecule as 16S rRNA. He and his team proceeded by extracting ribosomal RNA from the bacterial cells, purifying samples of the 16S molecules in each, and cutting those molecules into variously sized fragments with enzymes. Then they separated the fragments by electrophoresis, using an electrical field and a racetrack of soaked paper or gel.
In electrophoresis, a solution of mixed fragments is added to the racetrack, the power is turned on, and the electrical force pulls small fragments along faster than large ones, causing them to separate as distinct bands or ovals along the track. In Woese’s effort, each fragment comprised just a few of those A, C, G, U bases—maybe three, maybe five, maybe eight, maybe as many as twenty, but always a minuscule fraction of the full molecule. Those small fragments could then be pulled again, this time in a sideways direction, and their exact sequence would begin to come clear, based on the chemical and electrical differences among A, C, G, and U. Small fragments were easier to sequence by this method than one mammoth chain. AAG was easier to discern, as you might imagine, than AAUUUUUCAUUCG.
There were several stages of work. The primary run began the process of separating the fragments from one another. The secondary run, in a sideways dimension, revealed more about each fragment, which grew discretely recognizable as it raced not just down the racetrack but also now across. Those fragments, because of their radioactive content, showed as ovals burned onto the X-ray films. The oval-marked films would let an expert interpreter such as Woese infer the sequences—that