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Molecular Biology 1961

The Dependence of Cell-Free Protein Synthesis upon Synthetic Polyribonucleotides

Marshall Nirenberg & J. Heinrich Matthaei

Feed a cell a string of U's and it makes a protein of pure phenylalanine — the genetic code's first word, read aloud.

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In depth · the introduction

To read the secret code of life, two scientists fed a cell the simplest possible message — and listened to what it said back.

The big idea

Genes are written in a four-letter alphabet (the bases A, U, G, C in RNA), but living things are built from proteins, which are chains of 20 different amino acids. So there must be a code — a way of translating RNA letters into amino acids. Everyone believed the code was real, but in 1961 no one had read a single word of it.

Nirenberg and Matthaei found a way to read one. They wrote the simplest possible message themselves: a strand of RNA made of just one letter, U, repeated over and over. When they fed this 'poly-U' to the protein-making machinery of a cell, it churned out a protein made of just one amino acid — phenylalanine — over and over. The message 'UUU' meant 'phenylalanine.' The first word of the code had been read.

How it came about

By 1961 the coding problem was the great unsolved puzzle of the new molecular biology, and brilliant people had filled notebooks with clever theoretical codes — most of them wrong. Marshall Nirenberg, a young and little-known scientist at the U.S. National Institutes of Health, took a different route: an experiment. With the German biochemist Heinrich Matthaei, he built a 'cell-free' system — the working insides of broken-open bacteria, kept alive in a test tube and still able to make protein.

In the early hours of 27 May 1961, Matthaei ran the decisive tube: cell juice, the building blocks, and a synthetic strand of pure U. It produced a burst of phenylalanine protein. When Nirenberg announced it that summer at a congress in Moscow — at first to a nearly empty room — the news electrified the field. A code that had resisted the cleverest theorists had been cracked by a straightforward experiment.

Why it mattered

It turned the genetic code from an idea into something you could read, letter by letter. Once the method existed, the rest of the code fell quickly, giving us the universal dictionary that translates DNA into the proteins that build every living thing — the foundation of modern genetics, medicine and biotechnology.

A way to picture it

Imagine you have found a long coded message but have no key. So you cheat: you write your own message containing only the letter 'U', thousands of times, and slip it into the decoding machine. Out comes a translation that is only the word 'phenylalanine', thousands of times. Now you know one thing for certain — 'U' (in threes) means 'phenylalanine'. Do the same with a page of pure 'A', then pure 'C', and one entry at a time you rebuild the whole key. That is exactly how the code of life was first read.

An interactive diagram of a cell making protein. Choose a synthetic RNA made of one repeated letter — poly-U, poly-A, poly-C or poly-G — and slide how many codons are read. The RNA is split into groups of three, each group labelled with the amino acid it codes, and a chain of beads grows below to show the protein being built.

Where it sits

Watson and Crick (1953) showed DNA's structure; Crick's central dogma (1958) said information flows DNA → RNA → protein. But neither said how the letters translate. This experiment supplied the missing dictionary's first entry, and within a few years the whole code was known — the link between gene and protein that today's gene reading and editing all depend on.

The original document
Original source text
M. W. Nirenberg & J. H. Matthaei · PNAS 47 (1961): 1588–1602
The system
The paper works with a cell-free extract from Escherichia coli — broken-open cells that still supply ribosomes, transfer RNA, enzymes and an energy source — in which protein synthesis can be driven in a test tube. Adding an RNA template to this extract makes it incorporate radioactively labelled amino acids into newly made protein, which is then measured.
The synthetic templates
Instead of natural messenger RNA of unknown sequence, the authors added synthetic polyribonucleotides of known, simple composition — including polyuridylic acid (poly-U), a chain built from a single repeated base, U.
The decisive result
Poly-U specifically and dramatically stimulated the incorporation of one amino acid — phenylalanine — producing polyphenylalanine, while other amino acids were not. Because poly-U is nothing but a long run of U's, this tied a run of uridine to phenylalanine and, read as triplets, pointed to UUU as a codon for phenylalanine: the first entry of the genetic code to be deciphered.
What followed
The same approach soon assigned further codons — poly-A directing lysine, poly-C directing proline — and the complete 64-codon table was finished within a few years.
National Institutes of Health, Bethesda · 1961