Concept


In the 1920s, experiments showed that a harmless strain of bacteria can become infectious when mixed with a virulent strain of bacteria that had been killed. The dead bacteria apparently provide some chemical that "transforms" the harmless bacteria to infectious ones. This so-called "transforming principle" appeared to be a gene. A team of scientists led by Oswald Avery at the Rockefeller Institute, rigorously followed up on these experiments in the 1940's. They found that a pure extract of the "transforming principle" was unaffected by treatment with protein-digesting enzymes but was destroyed by a DNA-digesting enzyme. This showed that the transforming principle is DNA — and, by extension, a gene is made of DNA. Still, many scientists were slow to accept this clear proof that DNA, not protein, is the genetic molecule.

Animation


How do you do? I'm Oswald Avery. My colleagues and I did a series of experiments using strains of Pneumococcus bacteria, which cause pneumonia. Pneumoccous grows in the body of the host, but, like other types of bacteria, also can be grown on solid or liquid cultures. In 1928, Fred Griffith published a study on the different strains of Pneumoccous. Two in particular, S and R, look different. The S colonies have a smooth surface, and the R colonies look rough. The S colonies look smooth because each bacterium has a capsule-like coat made of sugars. This coat protects the S bacteria from the host's immune system, and so the S strain is infectious. The coat-less R strain is not. Griffith found that mice injected with the S strain develop pneumonia and die within days. Mice injected with the R strain do not get pneumonia. Griffith noticed that different strains of Pneumococcus could be cultured from one patient. He began to wonder if one strain could change into another. To test this idea, he did a series of experiments using the R and S strains. First, Griffith heated the S strain culture to kill the bacteria. As predicted, when injected into mice, the heat-killed bacteria did not produce an infection. Griffith co-injected the heat-killed S with live R into mice, and, much to his surprise, the mice developed pneumonia and died. Even more astonishing, Griffith was able to isolate live S strain from the blood of infected mice. These cultures could infect other mice. S strain cultured from infected mice remained active—showing that the change was stable and inherited. Griffith concluded that some "principle" was transferred from the heat-killed S to the R strain. The principle transformed the R into the infective S strain with a smooth coat. When I read about Griffith's results, I became very interested in the identity of this transforming "principle." Colin MacLeod, Maclyn McCarty, and I developed a test tube assay, so we no longer had to use mice to test for transformation. We used detergent to lyse the heat-killed S cells. Then we used this lysate for transformation assays. The test tube assays worked well, and showed us that the heat-killed S lysate could change R to S. The transforming principle was something in the lysate. We tested each of the lysate components for the transforming activity. First, we incubated the heat-killed S lysate with an enzyme, SIII, that completely chewed up the sugar coat. We tested the transforming ability of the sugar coat-less S lysates. The sugar coat-less S lysate was still able to transform. This told us that the R strain was not just assembling a new S sugar coat from the pieces. Next, we incubated the coat-less S extract with protein digesting enzymes — trypsin and chymotrypsin. We tested this lysate's ability to transform. This protein-less lysate was still able to transform. So, the transforming principle is not protein. While we were testing and purifying the lysate, we precipitated nucleic acids — DNA and RNA — with alcohol. We were the first to isolate nucleic acids from Pneumococcus. Since the transforming principle was not the sugar coat, and not protein, we suspected that it may be one of the nucleic acids. We dissolved the precipitate in water, and tested the transforming ability of the solution. First, we destroyed the RNA using the RNase enzyme. We tested this solution for its ability to transform. The solution still had the ability to transform. Therefore, RNA could not be the transforming principle. What we had left was virtually pure DNA. As a final test, we incubated the solution with the DNA-digesting enzyme, DNase. We used this solution to test for transforming ability. This solution was unable to transform. My colleagues and I concluded that DNA is the transforming principle, and we published these results in 1944.

Gallery


An Avery family portrait, 1886. Oswald is seated to the left of his father, the Reverend Joseph Francis Avery.
1900 picture of the Colgate band. Avery is seated in the middle holding his cornet.
Oswald Avery at work in the laboratory, around 1930.
Memo approving Avery's appointment to the Rockefeller Institute.
Avery at a 1940 Christmas party.
A page from the May 15, 1943 letter from Oswald Avery to his brother Roy. In the letter Avery speculated on how transformation could happen. Avery never publicly connected genes with DNA and his transformation experiments.
Colin Munro MacLeod, 1936.
Maclyn McCarty, 1936.

Audio/Video


Audio Glossary

Allele, Deoxyribonucleic acid (DNA), Gene, Genome, Genotype, Inherited, Phenotype

Video Interviews

Maclyn McCarty

Maclyn McCarty is Professor Emeritus at the Rockefeller University. He worked with Oswald Avery on studying and characterizing the transforming ability of DNA.

Clip 1 (1:00)
Commenting on Avery as a scientific group leader and as a person.

Clip 2 (0:53)
Relating how Avery was a successful orator while an undergraduate at Colgate University, and his subsequent disdain for public speaking as a scientist.

Clip 4 (0:40)
Describing the in vitro transformation experiments: the effect of removing polysaccharides from the bacterial extracts.

Clip 5 (0:37)
Describing the in vitro transformation experiments: the effect of destroying nucleic acids.

Clip 6 (0:26)
Characterizing the resistence to the discovery of DNA as the transforming factor: running against existing dogma.

Clip 7 (0:59)
How the bacterial transformation experiments provided the first real opportunity to study the chemical nature of the gene.

Biography


 

In 1944, Oswald Avery and his colleagues, Colin MacLeod and Maclyn McCarty published their landmark paper on the transforming ability of DNA.

OSWALD THEODORE AVERY (1877-1955)

Oswald Avery was born in 1877 in Halifax, Nova Scotia. His father was a Baptist minister, and when Oswald was ten, his father became the pastor at the Mariners' Temple in New York's Lower East Side. Avery's parents were strong, enterprising people. They managed quite well on a small pastoral salary in the midst of one of the most crowded and squalid areas of New York City. They received occasional monetary donations from John D. Rockefeller, the rich industrial, who was an active supporter of the Baptist Church.

As a young boy, Avery learned how to play the cornet and on Sunday afternoons would play to attract worshippers to the church. He was so good that he won a scholarship to the National Conservatory of Music. In 1893, Avery attended the Colgate Academy and later the Colgate University. He became the leader of the college band and acquired the nickname "Babe" because of his small stature.

Avery was a good student and graduated from Colgate with a B.A. Even though he took very few science courses at Colgate, after graduation Avery went to the College of Physicians and Surgeons to study medicine. At the turn of the century, the field of medicine was changing. Scientists were beginning to determine the nature and cause of disease. Though good with patients, Avery found medical research more intellectually satisfying.

In 1907, Avery accepted an associate directorship at Hoagland Laboratory in Brooklyn — the first privately endowed bacterial research laboratory in the United States. While at the Hoagland, Avery taught student nurses and earned another nickname: "Fess" for professor.

Avery worked on many strains of bacteria, applying different immunological and chemical methods. In 1913, Avery published a clinical study of the tuberculosis bacterium. This work attracted the attention of Dr. Rufus Cole, the director of the Rockefeller Institute Hospital, who offered Avery a job at the Rockefeller. Avery did his Pneumococcus work at the Rockefeller and stayed there until his retirement in 1948.

Avery was well-liked by his colleagues, even though he didn't spend much time socializing with them. He also traveled infrequently and rarely attended scientific conferences or meetings. The exception was his yearly summer vacation to Deer Island in Maine where he could indulge in one of his favorite pastimes, sailing.

After retirement, Avery moved to Nashville to be near his brother's family. Although he was offered an opportunity to continue his research career, Avery cultivated the lifestyle of the retired "country gentleman." He took long walks, gardened and spent time with his family. In 1954, he was diagnosed with liver cancer. Avery died the following year after a painful illness.

MACLYN MCCARTY (1911-)

Maclyn McCarty was born in South Bend, Indiana. His father worked for the Studebaker Corporation in South Bend and was sent to various locations around the country. His family moved with him and McCarty's early education was itinerant until they settled back in South Bend in 1922. His parents were both extremely well-read and they encouraged their children to be self-sufficient in the pursuit of knowledge.

In high school, McCarty read a book called Microbe Hunters. This led to other books on biology and medicine and helped McCarty decide on a career in medical research. As a prelude to later research, McCarty and three of his high school friends formed the "Amateur Research Chemists" Club and did experiments in their basement labs.

In 1929, McCarty went to Stanford University for premed training and in 1933 started medical school at Johns Hopkins University. He spent a few summers as a pediatrics intern but by the time he graduated medical school, McCarty was working in a clinical research lab.

After graduation, McCarty looked for a research position, a scarcity because of the war. In 1940, he accepted a position for $100 a month to work with William Tillet at New York University. The next year, McCarty obtained a fellowship from the National Research Council. Their recommendation was that McCarty use the fellowship in another lab to broaden his experimental horizons. Tillet made arrangements for McCarty to join Oswald Avery's lab at the Rockefeller Research Institute. Tillet had worked in Avery's lab in the 20's and thought that it was the natural place for McCarty to go to pursue his interest in bacteriological research.

At the Rockefeller, McCarty worked with Avery to perfect the purification of the Pneumococcus transforming factor, and they were the first to precipitate DNA from bacteria. They used enzymes to degrade different classes of molecules and proved that DNA was the transforming factor. Their landmark paper was published in 1944. McCarty had been drafted in 1942 and did most of the work in a naval uniform as part of the naval research unit based at Rockefeller Hospital.

In 1946, McCarty was given his own research lab at the Rockefeller Institute. He is now professor emeritus of the Rockefeller University, which evolved from the Rockefeller Research Institute.

Factoid

Links


 

Links

Profiles in Science

From the National Library of Medicine, this site has a very extensive collection of material on Oswald Avery. The material was donated by Dr. Joshua Lederberg.

Bibliography

  • Avery, Oswald, MacLeod, Colin, and McCarty, Maclyn, 1944, Studies on the Chemical Nature of the Substance Inducing Transformation of Pneumococcal Types, Journal of Exp. Med., 79: 137-158.

  • Bardossi, Fulvio, 1988, Oswald Avery and the Sugar-coated Microbe, Research Profiles, The Rockefeller University, 31, Spring.

  • Dubos, René J., 1976, The Professor, The Institute, and DNA, The Rockefeller University Press, New York.

  • Dunn, L.C., 1991, A Short History of Genetics: The Development of Some of the Main Lines of Thought: 1864-1939, Iowa State University Press, Ames.

  • Griffith, Fred, 1928, The Significance of Pneumococcal Types, Journal of Hygiene, Cambridge, 27: 113-159.

  • Judson, Horace Freeland, 1979, The Eighth Day of Creation: Makers of the Revolution in Biology, Simon and Schuster, New York.

  • Kay, Lily E., 1993, The Molecular Vision of Life, Oxford University Press, New York.

  • Lagerkvist, Ulf, 1998, DNA Pioneers and Their Legacy, Yale University Press, New Haven.

  • McCarty, Maclyn, 1985, The Transforming Principle: Discovering that Genes Are Made of DNA, W.W. Norton & Company, New York.

  • McCarty, Maclyn, 1994, A Retrospective Look: How We Identified the Pneumococcal Transforming Substance as DNA, Journal of Exp. Med., 179: 385-394.

  • Morange, Michel, 1998, A History of Molecular Biology, Harvard University Press, Cambridge, Massachusetts.

  • Nobel Lectures in Molecular Biology 1933-1975, 1977, Elsevier North-Holland, Inc., New York.

  • Olby, Robert, 1974, The Path to the Double Helix: The Discovery of DNA, Dover Publications, Inc., New York.

  • Portugal, Franklin H., and Cohen, Jack S., 1977, A Century of DNA: A History of the Structure and Function of the Genetic Substance, The Massachusetts Institute of Technology, Cambridge, Massachusetts.

  • Rosenfield, Israel, Ziff, Edward, and Van Loon, Borin, 1983, DNA for Beginners, Writers and Readers Publishing, Inc.

  • Watson, James D., 1987, Molecular Biology of the Gene, The Benjamin/Cummings Publishing Company, Inc., Menlo Park, California.

Glossary


Allele - One of the variant forms of a gene at a particular locus, or location, on a chromosome. Different alleles produce variation in inherited characteristics such as hair color or blood type. In an individual, one form of the allele (the dominant one) may be expressed more than another form (the recessive one).
Deoxyribonucleic acid (DNA) -
Gene - The functional and physical unit of heredity passed from parent to offpsring. Genes are pieces of DNA, and most genes contain the information for making a specific protein.
Genome - All the DNA contained in an organism or a cell, which includes both the chromosomes within the nucleus and the DNA in mitochondria.
Genotype - The genetic identity of an individual, which may not show as outward characteristics.
Inherited - Transmitted through genes from parents to offspring.
Phenotype - The observable traits or characteristics of an organism, for example hair color, weight, or the presence or absence of a disease. Phenotypic traits are not necessarily genetic.

Children resemble their parents.
Genes come in pairs.
Genes don't blend.
Some genes are dominant.
Genetic inheritance follows rules.
Genes are real things.
All cells arise from pre-existing cells.
Sex cells have one set of chromosomes; body cells have two.
Specialized chromosomes determine gender.
Chromosomes carry genes.
Genes get shuffled when chromosomes exchange pieces.
Evolution begins with the inheritance of gene variation.
Mendelian laws apply to human beings.
Mendelian genetics cannot fully explain human health and behavior.
DNA and proteins are the molecules of the cell nucleus.
One gene makes one protein.
Bacteria and viruses have DNA too.
The DNA molecule is shaped like a twisted ladder.
A half DNA ladder is a template for copying the whole.
RNA is an intermediary between DNA and protein.
DNA words are three letters long.
A gene is a discrete sequence of DNA nucleotides.
The RNA message is sometimes edited.
Some viruses store genetic information in RNA.
RNA was the first genetic molecule.
Mutations are changes in genetic information.
Some types of mutations are automatically repaired.
A chromosome is a package for DNA.
Higher cells incorporate an ancient chromosome.
Some DNA does not encode protein.
Some DNA can jump.
Genes can be turned on and off.
Genes can be moved between species.
DNA responds to signals from outside the cell.
Different genes are active in different kinds of cells.
Master genes control basic body plans.
Development balances cell growth and death.
A genome is an entire set of genes.
Living things share common genes.
DNA is only the starting point for understanding human biology.
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