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mRNA: History of Functional Investigation

By:Amy Ralston, Ph.D.(Write Science Right)&Kenna M. Shaw, Ph.D.(Nature
Education)2008Nature Education
Citation:Ralston,A.&Shaw,K.(2008)mRNA: history of functional
investigation.Nature Education1(1):124

When looking at a molecule that's transported around like RNA, how can you gure
out where it's synthesized and where it ends up? See how researchers examined
this very question in the 1950s.
During the mid1950s, the connection between DNA, RNA, and protein was still fuzzy. RNA had
been detected in both cytoplasm and nuclei, raising the question as to where this substance was
synthesized, and from what. For example, was RNA made in the nucleus from DNA and then
exported? Or was it made "from scratch" both inside and outside the nucleus by way of separate
processes? Resolving such questions was an essential rst step toward development of the proposal
that RNA is synthesized directly from DNA in the nucleus, then exported to the cytoplasm as a
messenger RNA (mRNA). But how exactly did researchers determine that RNA was of nuclear origin?

Observing RNA Synthesis

A simple way to determine where something comes from is to watch it arrive at its destination. In
the 1950s, however, live imaging of RNA synthesis presented a number of challenges. At that time,
researchers had developed techniques that allowed them to label RNA with radioactive phosphates,
which meant that they could visualize this RNA by preserving labeled samples in formaldehyde, then
putting the samples in contact with lm to produce autoradiographs. Using this method, the
researchers noticed that radioactivity rst appeared in cells' nuclei and only later in their cytoplasm.
Many scientists believed these ndings suggested that RNA must be made in the nucleus and then
transported out into the cytoplasm. In contrast, other scientists suggested that these images
actually meant that RNA was made in both locations, but merely at a faster rate in the nucleus. How,
then, could researchers determine whether RNA actually moved from the nucleus to the cytoplasm?

Goldstein and Plauts Experiments in Amoebas

Eventually, biologists Lester Goldstein and Walter Plaut devised a clever
strategy that allowed them to answer that very question through examination
of the relationship between nuclear and cytoplasmic RNA in amoebas.
Goldstein and Plaut began by labeling RNA with radioactivity using established
techniques. Next, the duo employed micromanipulation to carefully remove an
individual amoeba's nucleus and transfer it to an unlabeled, enucleated
amoeba. They reasoned that if RNA originated in the nucleus, radioactivity
would, at the end of the experiment, be detected in both nucleus and
cytoplasm. In contrast, if RNA did not trac from nucleus to cytoplasm, no
radioactivity would be detected in the cytoplasm at the conclusion of the
experiment.

Figures 1 &
2
Figure Detail

Goldstein and Plaut followed the voyage of the radioactive RNA by taking
samples at dierent times following the nuclear transfer, xing the samples, and exposing the xed
samples to lm to produce autoradiographs. Together, this series of images revealed the origin and
destination of nuclear RNA. Importantly, at early time points, essentially all the radioactivity was
detected within the nucleus (Figures 1A, B); then, at later time points, radioactivity was detected in
the cytoplasm as well (Figures 2A, B). The radioactivity had therefore moved from the nucleus to the
cytoplasm, strongly suggesting a nuclear origin for RNA.
The experiments of Goldstein and Plaut were consistent with the hypothesis that RNA is synthesized
in the nucleus and then exported to the cytoplasm. The experiment did not, however, rule out the

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in the nucleus and then exported to the cytoplasm. The experiment did not, however, rule out the
possibility that RNA is also made in the cytoplasm, although researchers do not currently believe
this to be the case. Can you think of a way to test this hypothesis?

References and Recommended Reading

Goldstein, L., & Plaut, W. Evidence for nuclear synthesis of cytoplasmic ribose nucleic acid. Proceedings of the National Academy of

Sciences 41, 874880 (1955).

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