RNA interference, is activated when RNA molecules occur as double-stranded pairs in the cell
THIS YEAR'S Nobel Laureates have discovered a fundamental mechanism for controlling the flow of genetic information.
Our genome operates by sending instructions for the manufacture of proteins from DNA in the nucleus of the cell to the protein synthesising machinery in the cytoplasm. These instructions are conveyed by messenger RNA (mRNA).
In 1998, the American scientists Andrew Fire and Craig Mello published their discovery of a mechanism that can degrade mRNA from a specific gene.
This mechanism, RNA interference, is activated when RNA molecules occur as double-stranded pairs in the cell. Double-stranded RNA activates biochemical machinery, which degrades those mRNA molecules that carry a genetic code identical to that of the double-stranded RNA.
When such mRNA molecules disappear, the corresponding gene is silenced and no protein of the encoded type is made.
RNA interference occurs in plants, animals, and humans. It is of great importance for the regulation of gene expression, participates in defence against viral infections, and keeps jumping genes under control. The genetic code in DNA determines how proteins are built. The instructions contained in the DNA are copied to mRNA and subsequently used to synthesize proteins . Our genome consists of approximately 30,000 genes.
However, only a fraction of them are used in each cell. Which genes are expressed (i.e. govern the synthesis of new proteins) is controlled by the machinery that copies DNA to mRNA in a process called transcription.
It, in turn, can be modulated by various factors. The fundamental principles for the regulation of gene expression were identified more than 40 years ago by the French Nobel Laureates François Jacob and Jacques Monod.
Today, we know that similar principles operate throughout evolution, from bacteria to humans.
They also form the basis for gene technology, in which a DNA sequence is introduced into a cell to produce new protein.
Andrew Z. Fire, Professor of Pathology and Genetics, Stanford University School of Medicine, U.S.
Around 1990, molecular biologists obtained a number of unexpected results that were difficult to explain. The most striking effects were observed by plant biologists who were trying to increase the colour intensity of the petals in petunias by introducing a gene inducing the formation of red pigment in the flowers.
But instead of intensifying the colour, this treatment led to a complete loss of colour and the petals turned white! The mechanism causing these effects remained enigmatic until Fire and Mello made the discovery for which they receive this year's Nobel Prize.
`Sense' and `antisense'
Andrew Fire and Craig Mello were investigating how gene expression is regulated in the nematode worm Caenorhabditis elegans . Injecting mRNA molecules encoding a muscle protein led to no changes in the behaviour of the worms.
The genetic code in mRNA is described as being the `sense' sequence, and injecting `antisense' RNA, which can pair with the mRNA, also had no effect.
But when Fire and Mello injected sense and antisense RNA together, they observed that the worms displayed peculiar, twitching movements. Similar movements were seen in worms that completely lacked a functioning gene for the muscle protein. When sense and antisense RNA molecules meet, they bind to each other and form double-stranded RNA.
Could it be that such a double-stranded RNA molecule silences the gene carrying the same code as this particular RNA?
Fire and Mello tested this hypothesis by injecting double-stranded RNA molecules containing the genetic codes for several other worm proteins. In every experiment, injection of double-stranded RNA carrying a genetic code led to silencing of the gene containing that particular code. The protein encoded by that gene was no longer formed.
Their discovery clarified many confusing and contradictory experimental observations and revealed a natural mechanism for controlling the flow of genetic information. The components of the RNAi machinery were identified during the following years.
Serves as a probe
Double-stranded RNA binds to a protein complex, Dicer, which cleaves it into fragments. Another protein complex, RISC, binds these fragments. One of the RNA strands is eliminated but the other remains bound to the RISC complex and serves as a probe to detect mRNA molecules.
When an mRNA molecule can pair with the RNA fragment on RISC, it is bound to the RISC complex, cleaved and degraded. The gene served by this particular mRNA has been silenced.
RNA interference is important in the defence against viruses, particularly in lower organisms. Many viruses have a genetic code that contains double-stranded RNA.
When such a virus infects a cell, it injects its RNA molecule, which immediately binds to Dicer.
The RISC complex is activated, viral RNA is degraded, and the cell survives the infection. Jumping genes, also known as transposons, are DNA sequences that can move around in the genome.
Craig C. Mello, Professor of Molecular Medicine and Howard Hughes Medical Institute Investigator, Program in Molecular Medicine, University of Massachusetts Medical School, U.S.
They are present in all organisms and can cause damage if they end up in the wrong place. Many transposons operate by copying their DNA to RNA, which is then reverse-transcribed back to DNA and inserted at another site in the genome.
Part of this RNA molecule is often double-stranded and can be targeted by RNA interference. In this way, RNA interference protects the genome against transposons.
New opportunities in biomedical research, gene technology and health care. RNA interference opens up exciting possibilities for use in gene technology.
Double-stranded RNA molecules have been designed to activate the silencing of specific genes in humans, animals or plants. Excerpted from public information available on http://nobelprize.org
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