The New Genetics by National Institutes of Health - HTML preview

PLEASE NOTE: This is an HTML preview only and some elements such as links or page numbers may be incorrect.
Download the book in PDF, ePub, Kindle for a complete version.

mistakenly believe that they are in the fly’s

so much that it can’t make proteins? Then the

thorax, and therefore ought to grow into a leg.

bacterium won’t grow, and Green wouldn’t find it.

And so they do.

Using clever molecular tricks, Green figured

Thinking about this odd situation taught

out a way to rescue some of the bacteria with

scientists an important lesson—that the proteins

defective ribosomes so they could grow. While

made by some genes can act as switches. Switch

some of the rescued bacteria have changes in

genes are master controllers that provide each

their ribosomal RNA that make them resistant

body part with a kind of identification card. If a

to certain antibiotics (and thus would not make

protein that normally instructs cells to become

good antibiotic targets) other RNA changes that

an antenna is disrupted, cells can receive new

don’t affect resistance may point to promising

instructions to become a leg instead.

ideas for new antibiotics.

index-25_1.jpg

index-25_2.jpg

The New Genetics I How Genes Work 21

FLYBASE; R. TURNER

. Normal fruit fly head.

. Fruit fly head showing the effects of the Antennapedia gene. This fly has legs where its antennae should be.

Scientists determined that several different

genes of different organisms, it’s a good clue

genes, each with a common sequence, provide

that these genes do something so important and

these anatomical identification card instructions.

useful that evolution uses the same sequence

Kaufman isolated and described one of these

over and over and permits very few changes in

genes, which became known as Antennapedia,

its structure as new species evolve.

a word that means “antenna feet.”

Researchers quickly discovered nearly

Kaufman then began looking a lot more

identical versions of homeobox DNA in almost

closely at the molecular structure of the

every nonbacterial cell they examined—from

Antennapedia gene. In the early 1980s, he and

yeast to plants, frogs, worms, beetles, chickens,

other researchers made a discovery that has been

mice and people.

fundamental to understanding evolution as well

Hundreds of homeoboxcontaining genes

as developmental biology.

have been identified, and the proteins they

The scientists found a short sequence of DNA,

make turn out to be involved in the early stages

now called the homeobox, that is present not only

of development of many species. For example,

in Antennapedia but in the several genes next to

researchers have found that abnormalities in

it and in genes in many other organisms. When

the homeobox genes can lead to extra fingers or

geneticists find very similar DNA sequences in the

toes in humans.

index-26_1.jpg

index-26_2.jpg

index-26_3.jpg

22 National Institute of General Medical Sciences The Tools of Genetics: Mighty Microarrays We now have the ability to attach a piece of every

but teachers and students are using them, too.

gene in a genome (all of an organism’s genes) to

The Genome Consortium for Active Teaching

a postage stampsized glass microscope slide.

program (www.bio.davidson.edu/GCAT) pro

This ordered series of DNA spots is called a DNA

vides resources and instructions for high school

microarray, a gene chip or a DNA chip.

and college students to do genechip experiments

Whichever name you prefer, the chip could

in class.

also be called revolutionary. This technology has

Microarrays are used to get clues about

changed the way many geneticists do their work

which genes are expressed to control cell, tissue

by making it possible to observe the activity of

or organ function. By measuring the level of RNA

thousands of genes at once.

production for every gene at the same time,

In recent years, microarrays have become

researchers can learn the genetic programming

standard equipment for modern biologists,

that makes cell types different and diseased cells

different from healthy ones.

The chips consist of large numbers of DNA

fragments distributed in rows in a very small

space. The arrays are laid out by robots that can

DNA fragments

DNA fragments are attached to

T The resulting pattern of fluorescence indicates

glass or plastic, then fluorescently

which genes are active.

tagged molecules are washed over

the fragments.

Complementary mRNA

Some molecules (green) bind to their

complementary sequence. These molecules can be identified because they glow under fluorescent light.

index-27_1.jpg

index-27_2.jpg

Got It?

Why are some infections hard

to treat with antibiotics? What

are some things researchers

might do to solve this public

health problem?

How does DNA work as a form

position DNA fragments so precisely that

In December 2004, the U.S. Food and

of information storage?

more than 20,000 of them can fit on one micro

Drug Administration cleared the first

scope slide.

gene chip for medical use. The Amplichip

Scientists isolate mRNA from cells grown

CYP450™, made by Roche Molecular Systems

How can 20,000 human genes

under two conditions and tag the two sources

Inc. of Pleasanton, California, analyzes variaprovide the instructions for of RNA with different colors of fluorescent moletions in two genes that play a major role in making hundreds of thousands

cules. The two colors of RNA are then placed

the body’s processing of many widely preof different proteins?

on the chip, where they attach to complementary

scribed drugs. This information can help

DNA fragments anchored to the chip’s surface.

doctors choose the proper dose of certain

Next, a scanner measures the amount of

medicines for an individual patient.

What newborn tests does your

fluorescence at each spot on the chip, revealing

area hospital routinely do?

how active each gene was (how much mRNA

each gene produced). A computer analyzes the

patterns of gene activity, providing a snapshot

of a genome under two conditions ( e.g. , healthy

or diseased).

index-28_1.jpg

C H A P T E R 2

RNA and DNA Revealed: New Roles, New Rules For many years, when scientists thought

about heredity, DNA was the first thing

to come to mind. It’s true that DNA is the basic

ingredient of our genes and, as such, it often

steals the limelight from RNA, the other form

C

G

of genetic material inside our cells.

U

C

But, while they are both types of genetic

Sugar

U

phosphate

material, RNA and DNA are rather different.

backbone

C

The chemical units of RNA are like those of

G

A

DNA, except that RNA has the nucleotide uracil

C

(U) instead of thymine (T). Unlike double

U

stranded DNA, RNA usually comes as only a single

C

G

strand. And the nucleotides in RNA contain ribose

G

sugar molecules in place of deoxyribose.

A

U

RNA is quite flexible—unlike DNA, which is

Base

U

a rigid, spiralstaircase molecule that is very stable.

G

RNA can twist itself into a variety of complicated,

C

U

threedimensional shapes. RNA is also unstable in

C

that cells constantly break it down and must continually make it fresh, while DNA is not broken C

A

down often. RNA’s instability lets cells change

G

their patterns of protein synthesis very quickly

C

A

in response to what’s going on around them.

C

Many textbooks still portray RNA as a passive

A

U

molecule, simply a “middle step” in the cell’s

Ribonucleic acid (RNA) has

genereading activities. But that view is no longer

the bases adenine (A),

cytosine (C), guanine (G)

accurate. Each year, researchers unlock new

and uracil (U).

secrets about RNA. These discoveries reveal that

it is truly a remarkable molecule and a multi talented actor in heredity.

RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA

index-29_1.jpg

The New Genetics I RNA and DNA Revealed: New Roles, New Rules 25

Riboswitches are RNA

sequences that control

gene activity. The riboswitch

shown here bends into a

special shape when it grips

tightly onto a molecule

called a metabolite (colored

BREAKER

balls) that bacteria need

to survive.

RONALD

Today, many scientists believe that RNA

because of its ability to lead a double life: to store

evolved on the Earth long before DNA did.

information and to conduct chemical reactions.

Researchers hypothesize — obviously, no one

In other words, in this world, RNA served the

was around to write this down — that RNA was

functions of both DNA and proteins.

a major participant in the chemical reactions

What does any of this have to do with human

that ultimately spawned the first signs of life

health? Plenty, it turns out.

on the planet.

Today’s researchers are harnessing some of

RNA’s flexibility and power. For example, through

RNA World

a strategy he calls directed evolution, molecular

At least two basic requirements exist for making

engineer Ronald R. Breaker of Yale University is

a cell: the ability to hook molecules together and

developing ways to create entirely new forms of

break them apart, and the ability to replicate, or

RNA and DNA that both work as enzymes.

copy itself, from existing information.

Breaker and others have also uncovered

RNA probably helped to form the first cell.

a hidden world of RNAs that play a major

The first organic molecules, meaning molecules

role in controlling gene activity, a job once

containing carbon, most likely arose out of random

thought to be performed exclusively by proteins.

collisions of gases in the Earth’s primitive atmos

These RNAs, which the scientists named

phere, energy from the Sun, and heat from naturally

riboswitches, are found in a wide variety of

occurring radioactivity. Some scientists think that

bacteria and other organisms.

in this primitive world, RNA was a critical molecule

RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA

index-30_1.jpg

index-30_2.jpg

index-30_3.jpg

index-30_4.jpg

26 National Institute of General Medical Sciences This discovery has led Breaker to speculate

that new kinds of antibiotic medicines could be

developed to target bacterial riboswitches.

Molecular Editor

Scientists are learning of another way to customize proteins: by RNA editing. Although DNA sequences spell out instructions for producing

RNA comes in a variety of

different shapes (above

RNA and proteins, these instructions aren’t

and right).

always followed precisely. Editing

a gene’s mRNA, even by a single

chemical letter, can radically change

Doublestranded DNA

the resulting protein’s function.

(left) is a staircaselike

molecule.

Nature likely evolved the RNA

editing function as a way to get more

proteins out of the same number of

Small But Powerful

Recently, molecules called microRNAs have been

found in organisms as diverse as plants, worms

Larger RNA

and people. The molecules are truly “micro,” consisting of only a few dozen nucleotides, compared to typical human mRNAs that are a few thousand

nucleotides long.

Dicer

enzyme

What’s

particularly interesting about microRNAs

is that many of them arise from DNA that used

to be considered merely filler material (see

MicroRNA

page 14).

How do these small but important RNA molecules do their work? They start out much bigger but get trimmed by cellular enzymes, including

mRNA

one aptly named Dicer. Like tiny pieces of

Nearperfect complementarity

to target mRNA

The enzyme Dicer generates microRNAs by

chopping larger RNA molecules into tiny

No translation

Velcro®like pieces. MicroRNAs stick to mRNA

molecules and prevent the mRNAs from being

made into proteins.

No protein

RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA

index-31_1.jpg

index-31_2.jpg

The New Genetics I RNA and DNA Revealed: New Roles, New Rules 27

genes. For example, researchers have found that

the RNA sequence, which in turn changes the

the mRNAs for certain proteins important for the

protein that gets made.

proper functioning of the nervous system are

Bass’ experiments show that RNA editing

particularly prone to editing. It may be that RNA

occurs in a variety of organisms, including peoediting gives certain brain cells the capacity to ple. Another interesting aspect of editing is that

react quickly to a changing environment.

certain diseasecausing microorganisms, such as

Which molecules serve as the editor and how

some forms of parasites, use RNA editing to gain

does this happen? Brenda Bass of the University of

a survival edge when living in a human host.

Utah School of Medicine in Salt Lake City studies

Understanding the details of this process is an

one particular class of editors called adenosine

important area of medical research.

deaminases. These enzymes “retype” RNA letters

at various places within an mRNA transcript.

They do their job by searching for characteristic RNA shapes. Telltale twists and bends in folded RNA molecules signal these enzymes to change

AM

Y

PASQUINELLI

Velcro®, microRNAs stick to certain mRNA molecules and stop them from passing on their Worms with a mutated form of the microRNA let7

proteinmaking instructions.

(right) have severe growth problems, rupturing as

First discovered in a roundworm model system

they develop.

(see Living Laboratories, page 49), some microRNAs help determine the organism’s body plan. In their

absence, very bad things can happen. For example, worms engineered to lack a microRNA called MicroRNA molecules also have been linked to

let7 develop so abnormally that they often rupture

cancer. For example, Gregory Hannon of the Cold

and practically break in half as the worm grows.

Spring Harbor Laboratory on Long Island, New

Perhaps it is not surprising that since microRNAs

York, found that certain microRNAs are associhelp specify the timing of an organism’s developated with the severity of the blood cancer Bcell mental plan, the appearance of the microRNAs

lymphoma in mice.

themselves is carefully timed inside a developing

Since the discovery of microRNAs in the

organism. Biologists, including Amy Pasquinelli

first years of the 21st century, scientists have

of the University of California, San Diego, are curidentified hundreds of them that likely exist as rently figuring out how microRNAs are made

part of a large family with similar nucleotide

and cut to size, as well as how they are produced

sequences. New roles for these molecules are

at the proper time during development.

still being found.

RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA 28 National Institute of General Medical Sciences RNA Interference (RNAi)

Dicer enzyme

Doublestranded RNA (dsRNA) is chopped

dsRNA

into short interfering RNAs (siRNAs) by the

enzyme Dicer.

Short interfering

RNAs (siRNAs)

A

G U A

G

U

C

C

RISC

A

G U

G

The RNAinduced silencing

U C

A C

complex (RISC) enzyme

U

C A

C

A G

U G

attaches to siRNA.

mRNA

The siRNARISC complex

attaches to target mRNA

and chops the mRNA into

small pieces.

Chopped mRNA

(no longer functional)

RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA RNA

index-33_1.jpg

index-33_2.jpg

The New Genetics I RNA and DNA Revealed: New Roles, New Rules 29

Healthy Interference

RNA controls genes in a way that was only discovered recently: a process called RNA interference, or RNAi. Although scientists identified RNAi less

than 10 years ago, they now know that organisms

have been using this trick for millions of years.

Researchers believe that RNAi arose as a way to

of genes that affect cell growth and tissue

reduce the production of a gene’s encoded protein

formation in roundworms, using a molecular

for purposes of finetuning growth or selfdefense.

tool called antisense RNA.

When viruses infect cells, for example, they com

To their surprise, Mello and Fire found

mand their host to produce specialized RNAs

that their antisense RNA tool wasn’t doing

that allow the virus to survive and make copies

much at all. Rather, they determined, a doubleof itself. Researchers believe that RNAi eliminates stranded contaminant produced during the

unwanted viral RNA, and some speculate that

synthesis of the singlestranded antisense RNA

it may even play a role in human immunity.

interfered with gene expression. Mello and

Oddly enough, scientists discovered RNAi

Fire named the process RNAi, and in 2006 were

from a failed experiment! Researchers investiawarded the Nobel Prize in physiology or gating genes involved in plant growth noticed

medicine for their discovery.

something strange: When they tried to turn

Further experiments revealed that the doublepetunia flowers purple by adding an extra stranded RNA gets chopped up inside the cell

“purple” gene, the flowers bloomed white instead.

into much smaller pieces that stick to mRNA and

This result fascinated researchers, who could

block its action, much like the microRNA pieces

not understand how adding genetic material

of Velcro discussed above (see drawing, page 28).

could somehow get rid of an inherited trait. The

Today, scientists are taking a cue from nature

mystery remained unsolved until, a few years

and using RNAi to explore biology. They have

later, two geneticists studying development saw

learned, for example, that the process is not limited

a similar thing happening in lab animals.

to worms