Transformation
Griffith’s Experiments
• In 1928, Frederick Griffith, a bacteriologist, was trying to prepare a vaccine against pneumonia.
• A vaccine is a substance that is prepared from killed or weakened disease-causing agents, including certain
bacteria.
• The vaccine is introduced into the body to protect the body against future infections by the disease-causing
agent.
• Griffith discovered that harmless bacteria could turn virulent when mixed with bacteria that cause disease.
• A bacteria that is virulent is able to cause disease.
• Griffith had discovered what is now called transformation, a change in genotype caused when cells take up
foreign genetic material.
Avery’s Experiments
• In 1944, a series of experiments showed that the activity of the material responsible for transformation is not
affected by protein-destroying enzymes. The activity is stopped, however, by a DNA-destroying enzyme.
• Thus, almost 100 years after Mendel’s experiments, Oswald Avery and his co-workers demonstrated that DNA is
the material responsible for transformation.
DNA’s Role Revealed
• In 1952, Alfred Hershey and Martha Chase used the bacteriophage T2 to prove that DNA carried genetic
material.
• A bacteriophage, also referred to as phage, is a virus that infects bacteria.
When phages infect bacterial cells, the phages are able to produce more viruses, which are released when the
bacterial cells rupture
DNA’s Role Revealed
• Hershey and Chase carried out the following experiment:
Step 1 T2 phages were labeled with radioactive isotopes.
Step 2 The phages infect E. coli bacterial cells.
Step 3 Bacterial cells were spun to remove the virus's protein coats.
• Hershey and Chase concluded that the DNA of viruses is injected into the bacterial cells, while most of the viral
proteins remain outside.
- The injected DNA molecules causes the bacterial cells to produce more viral DNA and proteins
• This meant that the DNA, rather than proteins, is the hereditary material, at least in viruses.
A Winding Staircase
• Watson and Crick determined that a DNA molecule is a double helix—two strands twisted around each other,
like a winding staircase.
• Nucleotides are the subunits that make up DNA. Each nucleotide is made of three parts: a phosphate group, a
five-carbon sugar molecule, and a nitrogen-containing base.
• The five-carbon sugar in DNA nucleotides is called deoxyribose.
The nitrogen base in a nucleotide can be either a bulky, double-ring purine, or a smaller, single-ring pyrimidine
Chargaff’s Observations
• In 1949, Erwin Chargaff observed that for each organism he studied, the amount of adenine always equaled the
amount of thymine (A=T).
• Likewise, the amount of guanine always equaled the amount of cytosine (G=C).
• However, the amount of adenine and thymine and of guanine and cytosine varied between different organisms.
Wilkins and Franklin’s Photographs
• By analyzing the complex patterns on X-ray diffraction photo, scientists can determine the structure of the
molecule.
• In 1952, Maurice Wilkins and Rosalind Franklin developed high-quality X-ray diffraction photographs of strands
of DNA.
These photographs suggested that the DNA molecule resembled a tightly coiled helix and was composed of two
or three chains of nucleotides
Watson and Crick’s DNA Model
• In 1953, Watson and Crick built a model of DNA with the configuration of a double helix, a “spiral staircase” of
two strands of nucleotides twisting around a central axis.
The double-helical model of DNA takes into account Chargaff’s observations and the patterns on Franklin’s X-ray
diffraction photographs
Pairing Between Bases
• An adenine on one strand always pairs with a thymine on the opposite strand, and a guanine on one strand
always pairs with a cytosine on the opposite strand.
• These base-pairing rules are supported by Chargaff’s observations.
The strictness of base-pairing results in two strands that contain complementary base pairs
• The diagram of DNA below the helix makes it easier to visualize the base-pairing that occurs between DNA
strands.
Roles of Enzymes in DNA Replication
• The complementary structure of DNA is used as a basis to make exact copies of the DNA each time a cell divided.
• The process of making a copy of DNA is called DNA replication.
• DNA replication occurs during the synthesis (S) phase of the cell cycle, before a cell divides.
• DNA replication occurs in three steps:
Step 1 DNA helicases open the double helix by breaking the hydrogen bonds that link the complementary
nitrogen bases between the two strands. The areas where the double helix separates are called replication
forks.
Step 2 At the replication fork, enzymes known as DNA polymerases move along each of the DNA strands. DNA
polymerases add nucleotides to the exposed nitrogen bases, according to the base-pairing rules.
Step 3 Two DNA molecules form that are identical to the original DNA molecule
Checking for Errors
• In the course of DNA replication, errors sometimes occur and the wrong nucleotide is added to the new strand.
• An important feature of DNA replication is that DNA polymerases have a “proofreading” role.
• This proofreading reduces errors in DNA replication to about one error per 1 billion nucleotides.
The Rate of Replication
• Replication does not begin at one end of the DNA molecule and end at the other.
• The circular DNA molecules found in prokaryotes usually have two replication forks that begin at a single point.
• The replication forks move away from each other until they meet on the opposite side of the DNA circle.
• In eukaryotic cells, each chromosome contains a single, long strand of DNA.
• Each human chromosome is replicated in about 100 sections that are 100,000 nucleotides long, each section
with its own starting point.
• With multiple replication forks working in concert, an entire human chromosome can be replicated in about 8
hours.
Decoding the Information in DNA
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Traits, such as eye color, are determined by proteins that are built according to instructions coded in DNA.
Proteins, however, are not built directly from DNA. Ribonucleic acid is also involved.
Like DNA, ribonucleic acid (RNA) is a nucleic acid—a molecule made of nucleotides linked together.
RNA differs from DNA in three ways:
1. RNA consists of a single strand of nucleotides instead of the two strands found in DNA.
2. RNA nucleotides contain the five-carbon sugar ribose rather than the sugar deoxyribose, which is found in
DNA nucleotides.
3. In addition to the A, G, and C nitrogen bases found in DNA, RNA nucleotides can have a nitrogen base called
uracil (U).
• The instructions for making a protein are transferred from a gene to an RNA molecule in a process called
transcription.
• Cells then use two different types of RNA to read the instructions on the RNA molecule and put together the
amino acids that make up the protein in a process called translation.
• The entire process by which proteins are made based on the information encoded in DNA is called gene
expression, or protein synthesis.
Transfer of Information from DNA to RNA
• The first step in the making of a protein, transcription, takes the information found in a gene in the DNA and
transfers it to a molecule of RNA.
• RNA polymerase, an enzyme that adds and links complementary RNA nucleotides during transcription, is
required.
• The three steps of transcription are:
Step 1 RNA polymerase binds to the gene’s promoter.
Step 2 The two DNA strands unwind and separate.
Step 3 Complementary RNA nucleotides are added
The Genetic Code: Three-Nucleotide “Words
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Different types of RNA are made during transcription, depending on the gene being expressed.
When a cell needs a particular protein, it is messenger RNA that is made.
Messenger RNA (mRNA) is a form of RNA that carries the instructions for making a protein from a gene and
delivers it to the site of translation.
• The information is translated from the language of RNA—nucleotides—to the language of proteins—amino
acids.
• The RNA instructions are written as a series of three-nucleotide sequences on the mRNA called codons.
• The genetic code of mRNA is the amino acids and “start” and “stop” signals that are coded for by each of the
possible 64 mRNA codons.
RNA’s Roles in Translation
• Translation takes place in the cytoplasm. Here transfer RNA molecules and ribosomes help in the synthesis of
proteins.
• Transfer RNA (tRNA) molecules are single strands of RNA that temporarily carry a specific amino acid on one
end.
• An anticodon is a three-nucleotide sequence on a tRNA that is complementary to an mRNA codon.
• Ribosomes are composed of both proteins and ribosomal RNA (rRNA).
• Ribosomal RNA (rRNA) molecules are RNA molecules that are part of the structure of ribosomes.
• Each ribosome temporarily holds one mRNA and two tRNA molecules.
• The seven steps of translation are:
Step 1 The ribosomal subunits, the mRNA, and the tRNA carrying methionine bind together.
Step 2 The tRNA carrying the amino acid specified by the codon in the A site arrives.
Step 3 A peptide bond forms between adjacent amino acids.
Step 4 The tRNA in the P site detaches and leaves its amino acid behind.
Step 5 The tRNA in the A site moves to the P site. The tRNA carrying the amino acid specified by the codon in the
A site arrives.
Step 6 A peptide bond is formed. The tRNA in the P site detaches and leaves its amino acid behind.
Step 7 The process is repeated until a stop codon is reached. The ribosome complex falls apart. The newly made
protein is released.
Protein Synthesis in Prokaryotes
• Both prokaryotic and eukaryotic cells are able to regulate which genes are expressed and which are not,
depending on the cell’s needs.
• The piece of DNA that overlaps the promoter site and serves as the on-off switch is called an operator.
• In bacteria, a group of genes that code for enzymes involved in the same function, their promoter site, and the
operator that controls them all function together as an operon.
• The operon that controls the metabolism of lactose is called the lac operon.
• When there is no lactose in the bacterial cell, a repressor turns the operon off.
• A repressor is a protein that binds to an operator and physically blocks RNA polymerase from binding to a
promoter site.
Protein Synthesis in Eukaryotes
Controlling the Onset of Transcription
• Most gene regulation in eukaryotes controls the onset of transcription—when RNA polymerase binds to a gene.
• Transcription factors help arrange RNA polymerases in the correct position on the promoter.
• An enhancer is a sequence of DNA that can be bound by a transcription factor.
Intervening DNA in Eukaryotic Genes
• In eukaryotes, many genes are interrupted by introns—long segments of nucleotides that have no coding
information.
• Exons are the portions of a gene that are translated (expressed) into proteins.
After a eukaryotic gene is transcribed, the introns in the resulting mRNA are cut out by complex assemblies of RNA
and protein called spliceosomes
Mutations
• Mutations that move an entire gene to a new location are called gene rearrangements.
• Changes in a gene’s position often disrupt the gene’s function because the gene is exposed to new regulatory
controls in its new location.
Genes sometimes move as part of a transposon. Other times, the portion of the chromosome containing a gene may
be rearranged during meiosis
• Mutations that change a gene are called gene alterations.
• In a point mutation, a single nucleotide changes.
• In an insertion mutation, a sizable length of DNA is inserted into a gene.
• In a deletion mutation, segments of a gene are lost, often during meiosis.
• Because the genetic message is read as a series of triplet nucleotides, insertions and deletions of one or two
nucleotides can upset the triplet groupings.
• A mutation that causes a gene to be read in the wrong three-nucleotide sequence is called a frameshift
mutation.