Macleod, Avery, McCarty: Inducing chemical changes in pneumococcus bacteria

Context:
Before this paper was published, the gene was "an entity without substance" The chemical identity
of the gene was unknown until Avery/McCarty/MacLeod published their paper in 1944 which
showed for the first time that DNA was the carrier of genetic material.

This experiment was based on the results of
Griffiths experiments:
Pneumococcus is the bacteria that causes pneumonia.
The two strains used in this study are the avirulent
(non pathogenic) R strain and the virulent (pathogenic)
S strain.
The S strain is classified based on the antigenic
polysaccharide capsule that induces an immunological
response in the host cell
The host immune system can only protect itself when
there is no capsule.
4 mice were injected with different bacteria cocktails
The results can be seen in the Figure 1

Griffiths Conclusions:
R-strain developed a capsule when it absorbed genetic material released from S-strain
There must be some "transforming principle" in living cells that is inducing chemical
changes in the R cells
Important because, until Frederick Griffith's studies, bacteriologists believed that different
serological types of pneumococcus were fixed and unchangeable from one generation to
the next.

Avery/McCarty/MacLeods Experiments:
The bacterial filtrate from the mouse killed by the heat killed S strain + R strain was extracted, this
filtrate contained the transforming principle. Various tests were run on this substance:
Test 1:
1. Deoxyribonuclease (DNAse)
which is an enzyme that
depolymerizes (breaks
down) DNA is naturally
present in various animal
serums
2. The serums were heated at
different temperatures, and
the control serum was not
heated
3. The R-Type Strain was then
grown in the serum with the
extracted transforming
principle
4. The idea was to test if the
presence of functional DNAse
inhibited transformation
Figure 1
Figure 2

Result: S colonies grew in the control serum (no DNase) and in the heated animal serums
(denatured DNase). Growth of S colonies indicates transformation occurred. Implies that DNase
inhibits transformation and DNA induces transformation.

Test 2: Chemical analysis showed that the ratio of nitrogen atoms to phosphorous atoms was
approximately 1.67 to 1, this is very close to the actual ratio of N:P in DNA

Test 3: Digestive enzyme tests: protease and RNAse had no effect on transformation. See Figure 2

Test 4: Conducted to find the minimum concentration of transforming principle to induce
transformation. The required amount was very small, and its concentration actually rose after
transformed cells reproduced, suggesting that the transformed cells actually began to synthesize it
themselves. This increase in concentration corroborates the hypothesis of DNA as a carrier of
heritable traits

Conclusions:
The transforming principle was a polymerized form of nucleic acid
Since there was no inherent biochemical specificity in nucleic acids like there is in antigens,
they were hesitant to conclude that DNA was the sole carrier of genetic information until
further experiments were conducted
"a nucleic acid of the desoxyribose type is the fundamental unit of the transforming
principle of Type III pneumococcus" (quote from actual paper)
Transformation can best be observed in the exponential growth phase of a bacterial colony
rather than in resting cells

Significance:
Perceived differences in prokaryotic and eukaryotic reproduction and the the fact that
transformation only occurred in bacteria caused the experiment to be viewed only as
medically significant.
Later work by Chargaff showed that the proportions of different nucleotides were not fixed
between species, adding support to the idea that, viewed as a polymer rather than
individual nucleic acids, DNA could in fact have the specificity to carry genetic information.
Until Watson, Crick, and Chargaff's experiments, the experiment was largely ignored by the
scientific community.