Re: GAL4/UAS in Drosophilia

Melanogaster

Postby SciB Sat Oct 10, 2015 8:47 pm

Hi,

I noticed that no one has answered your question, so I

thought I would give it a shot. The Gal4/UAS system is a
great way to visualize gene expression in a variety of
organisms including fruit flies. As you said, it is a little hard
to understand if you haven't had molecular biology.

The first thing you need to know is what the two acronyms,
Gal4 and UAS, stand for. Both of them are proteins, which
means they are encoded in genes in the DNA. They are
actually from bread yeast, but they work the same way in
Drosophila.

Gal4 is an activator of gene transcription, the synthesis of

messenger RNA (mRNA) from DNA. The 'Gal' part comes from
the name of a sugar, galactose. The Gal transcription
factors were first discovered in yeast as the activators of
galactose metabolism. You don't really need to know that
but I put it in so you won't be wondering where the name
comes from.

Transcription factors work by binding to a specific DNA

sequence that is ahead [the correct term is 'upstream'] of a
particular gene. When they are bound the gene can be
transcribed and the resulting mRNA used to make the
protein that is encoded by that gene.

The acronym UAS stands for 'upstream activation sequence',

and guess what--that is the sequence Gal4 binds to!

OK. So what is Gal4 and the UAS good for? This is where it
gets a little complicated so if you have more questions
please post again. Molecular biologists who study the
expression of specific genes, like for example some genes
that are switched on or off in Parkinson's disease, want to
have what is known as a reporter so they have a visual sign
of when the gene is being transcribed. Most genes have an
upstream sequence known as a promoter that sits right
before the transcription start site and activates
transcription.

What Drosophila researchers have done over the years is to

create a bunch (probably in the 1000s by now) of mutant
fruit flies that have the Gal4 gene placed downstream
(towards the transcription start site) of the promoter of a
certain gene that they are studying. It can be any gene. The
Gal4 sequence is engineered into the Drosophila genome and
when the flies reproduce, the offspring all have the Gal4
mutation in the same cells.

Now for the next part of the story. So, you have a fruit fly
that has a Gal4 gene just after the promoter of the gene you
are studying and you want to know when it is expressed.
How do you do that? Remember that the Gal4 protein is
itself an activator of gene transcription, but it needs its UAS
to bind to in order to work. This is where the reporter
comes in, and this is the complicated part, although once
you get the concept it is rather simple and elegant.

A reporter is a gene for a protein that can be visualized in

some way. A very common one is called green-fluorescent
protein (GFP) and as its name tells you, it gives of a green
glow when it is illuminated by light of a certain wavelength.
If you google 'reporter genes' you can find out more about
these important tools if you are interested.

The GFP gene actually came from a jellyfish so it is not

normally found in Drosophila; but by putting the GFP gene
under the control of a UAS, whenever Gal4 is produced, it
will bind to the UAS, activate transcription of GFP and the
cells in which it is present will glow green under a
fluorescence microscope. That tells you that the promoter
that was upstream of Gal4 in the study gene was activated.
Now you are getting some information about what might
control the expression of specific genes that might be
associated with the onset or progression of Parkinson's
disease.

This was a long explanation and I hope you understand now

how Gal4/UAS is used as a gene reporter system. It is all
done by molecular biology and genetic engineering and a lot
of hours spent in the lab by a lot of dedicated scientists.
Knowing which genes are expressed at which times in which
cells is the most important thing in biology. Figuring out the
sequence of the human genome was a great
accomplishment, but the real job is to know how all those
genes work in health and disease. Maybe you will be the one
to discover a new gene expression system in Parkinson's
disease that can be used to stop or prevent it.

If you still have questions, please don't hesitate to ask.

That's what we are here for.

Sybee