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Biology 1977

Phylogenetic Structure of the Prokaryotic Domain: The Primary Kingdoms

Carl R. Woese & George E. Fox

Reading life's oldest molecule, they found not two kinds of cell but three — and a whole hidden kingdom.

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In depth · the introduction

Looking for the deepest branches on the tree of life, two scientists found one nobody had ever drawn — a third kind of life, hiding in plain sight.

Not two kinds of life, but three

For a long time biology sorted all life into two boxes: the simple cells without a nucleus (bacteria) and the complex cells with one (everything else — us, plants, fungi). Carl Woese suspected this missed something, because it sorted cells by how they look, not by how they are related.

So he picked a molecule every living cell carries: ribosomal RNA, part of the tiny machine that builds proteins. By comparing its sequence between organisms, he could measure kinship directly. And out of that comparison fell a shock — a group of methane-making microbes that looked like bacteria but were, deep down, no more bacterial than they were like us. They were a whole third branch of life. Today we call them the Archaea.

The branch nobody believed in

Woese, working at the University of Illinois with his colleague George Fox, spent years on painstaking lab work: cutting up ribosomal RNA, reading its fragments, and building a 'catalog' for each organism. It was slow, unglamorous, and far from the famous problems of the day.

When the result came in 1977 — that the methane-makers were a separate, ancient lineage — much of biology simply did not buy it. Senior scientists were openly skeptical; Woese was dismissed by some as a crank for redrawing the tree of life on the strength of one molecule. He was right. As DNA sequencing matured, the third branch held, again and again, and the Archaea became textbook fact.

Why it matters

Most of life is microbial and invisible, and most microbes have never been grown in a laboratory — so for centuries we were blind to the true shape of life. Woese's method gave us eyes: read one universal molecule, and you can place any organism on the tree, even one you can only find as a smear of DNA in seawater or soil.

That changed what we could even ask. The Archaea turned out to be everywhere, including inside you, and they sit right next to the origin of complex cells — the question of where our own kind of cell came from. The map of life we use today is the one Woese and Fox started drawing.

Like comparing a shared sentence

Imagine every living thing carries the same very old paragraph, copied and recopied for billions of years. No copy is perfect, so small errors pile up — and the more two paragraphs differ, the longer ago they shared a copyist. Read those paragraphs side by side and you can rebuild the family tree, even for relatives you never knew existed.

Ribosomal RNA is that shared paragraph. When Woese lined up the methane-makers' copy against a bacterium's, the differences were as large as between a bacterium's and our own — telling him these were not bacteria at all, but cousins as distant as cousins get.

A similarity grid and six organisms. Sliding a cut-off joins close relatives; the organisms settle into three groups — two bacteria, two methane-makers, two eukaryotes — that stay separate across most settings.

Where it sits

Darwin (1859) gave life a single branching tree but no way to read its deepest forks; the discovery that genes are made of DNA, and the molecular biology that followed, finally made those forks readable. Woese and Fox supplied the molecule and the method. Read alongside the Margulis entry in this Library, the picture closes: our own complex cells appear to be a partnership between an archaeal host and a captured bacterium — so the three-domain tree and the cell-within-a-cell are two halves of one origin story.

The original document
Original source text
Carl R. Woese & George E. Fox · Proc. Natl. Acad. Sci. USA · vol. 74, no. 11, pp. 5088–5090 · November 1977
Abstract
A phylogenetic analysis based upon ribosomal RNA sequence characterization reveals that living systems represent one of three aboriginal lines of descent: (i) the eubacteria, comprising all typical bacteria; (ii) the archaebacteria, containing methanogenic bacteria; and (iii) the urkaryotes, now represented in the cytoplasmic component of eukaryotic cells.
The method, in brief
For each organism the small-subunit ribosomal RNA (16S, or 18S in eukaryotes) was cut into pieces with the enzyme RNase T1 and the resulting oligonucleotides were sequenced, giving a “catalog” of short sequences for that organism. Any two catalogs were then compared by a single similarity number, the binary association coefficient S_AB = 2N_AB/(N_A + N_B), where N_A and N_B count the nucleotides in each catalog's hexamer-and-larger oligomers and N_AB those in the oligomers the two share. The whole paper rests on one small table of these coefficients.
The three primary kingdoms
The methanogens — bacteria that make methane and that until then had been filed among ordinary bacteria — turned out to score no closer to typical bacteria than to the cytoplasm of eukaryotes: about the background value shared by all life. They were therefore not bacteria at all, but a separate, equally ancient lineage the authors named the archaebacteria. Life was thus divided into three “primary kingdoms” (urkingdoms): eubacteria, archaebacteria, and the urkaryote line preserved in the eukaryotic cytoplasm.
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Department of Genetics and Development, University of Illinois, Urbana · 1977