Lesson 7:

From Plan to
Protein
BIOL 1345
 Learning Objectives (Study Guide)
By the end of this lesson, students will be able to:
1. Explain the main differences between DNA, RNA, & proteins.
2. Identify the locations of DNA & the various RNAs in the cell.
3. Explain the relationship between DNA, a gene, nucleotides, amino acids, and proteins.
4. List the parts of a gene & describe the functions of each.
5. Describe how the process of transcription works.
6. Transcribe a DNA sequence into an mRNA sequence.
7. Describe the structures that are added to & removed from an mRNA transcript before
it leaves the nucleus.
8. Explain what alternative splicing is & why it is important for cells.
9. Identify the organelles that perform the process of translation.
 Learning Objectives (Study Guide)
By the end of this lesson, students will be able to:
10.Describe how the process of translation works.
11.Explain what a codon is.
12.Use the Genetic Code Table to translate an mRNA sequence.
13.Explain what differential gene expression is & why it is used in cells.
14.Describe the role of transcription factors in differential gene expression.
15.Explain what occurs in a point mutation & a frameshift mutation.
16.Describe the effects of silent mutations, missense mutations, and nonsense
mutations on translation.
17.Identify the major sources of mutations in a cell.
18.Describe how a cell responds to mutations.
 Remember, You are Your own
Instruction Manual!

In the last lecture we discussed how

DNA makes more DNA (DNA
replication).

In this lecture we will learn about

how cells assemble proteins, using
the instructions in the DNA.
 Gene Expression
Gene 2
Gene expression is the process of using DNA
molecule
the information stored in a gene to build a Gene 1

functional protein Gene 3

A gene is a region of DNA that carries the

instructions for making specific proteins. DNA
template
strand

Gene expression involves 2 steps:

• Step #1 – Transcription TRANSCRIPTION

• DNA for a specific gene is copied into mRNA

mRNA
Codon
• Step #2 – Translation TRANSLATION
• mRNA is used as a template for assembling a
protein Protein

Amino acid
 Gene Expression and RNA
There are multiple kinds of RNA involved in protein synthesis:
• Messenger RNA (mRNA) carries protein-building information from
DNA to the ribosome
• Ribosomal RNA (rRNA) makes up ribosomes
• Transfer RNA (tRNA) brings amino acids to the ribosomes
 Transcription
• During transcription, mRNA is synthesized from a DNA template. RNA contains
uracil in place of thymine.
• DNA-RNA base pairing follows these rules:
• A-U
• C-G
• mRNA then carries the instructions out of the nucleus to the ribosome
 Reminder: DNA vs. mRNA
Transcription uses the DNA template to build mRNA.

DNA is double- mRNA is single-

stranded stranded

DNA is made with mRNA is made with

deoxyribose sugar ribose sugar

DNA includes A, T, mRNA includes A, U,

G, & C nucleotides G, & C nucleotides
DNA is found in the
cell’s nucleus mRNA leaves the
nucleus & enters the
cytoplasm
 Let’s Practice!

Use the DNA strand below to transcribe the complementary mRNA strand:

3’ T A C G G T A G C G A T T T C 5’
 Stop & Think it Through!
DNA mRNA

Where is it found?

What sugar does it contain?

List its 4 nucleotides

Is it typically single-stranded
or double-stranded?

What is the function of this

molecule?
 The Central Dogma:
An Overview
Transcription occurs in the nucleus of a
eukaryotic cell
• RNA polymerase opens the DNA double helix &
creates a complementary pre-mRNA strand using
the sequence of nucleotides in a gene
• That pre-mRNA is processed, then leaves the
nucleus

Translation occurs in the cytoplasm of a

eukaryotic cell
• The mRNA molecule attaches to a ribosome
• The ribosome “reads” the mRNA & creates a
complementary amino acid chain
 Parts of a Gene
The promoter region of a gene is the The terminator region (a.k.a. stop sequence)
location where RNA Polymerase signals to RNA Polymerase that the whole gene
attaches to the DNA molecule. has been transcribed, so it can release the DNA.

The specific directions for

building the protein are stored
in this part of the gene.
Transcription: An Overview
Step 1 – Initiation
• RNA Polymerase attaches to the
DNA strand at the promoter
• Transcription factors are proteins
that bind to the DNA and regulate
transcription
Step 2 – Elongation
• RNA Polymerase creates a
complementary pre-mRNA molecule
Step 3 – Termination
• RNA Polymerase releases the pre-
mRNA & DNA
 Introns vs. Exons
Pre-mRNA molecules include introns & exons
Introns are “intervening sequences”
between the exons in a pre-mRNA
molecule
• Introns do NOT have information
involved in protein synthesis

Exons are the protein-coding

regions of a pre-mRNA molecule
• After the introns are spliced out,
these exons (“coding regions”) are
spliced together
 Alternative Splicing
• Alternative splicing allows a cell to make more than one protein using
a single gene
• Exons are selectively removed from a pre-mRNA transcript
• This can generate multiple different mRNA molecules from the same pre-mRNA
transcript
 Translation

• mRNA is used by the

ribosome as the instructions
for assembling amino acids
in the correct order
• tRNA (transfer RNA) carries
amino acids to the ribosome
so they can be assembled
into a protein Each tRNA molecule has an:
• amino acid attachment site that enables tRNA to
bring amino acids to the ribosome
• anticodon, the 3 nucleotides it will use to “read”
the mRNA codons
 Ribosome Structure
Ribosomes are made of ribosomal RNA
(rRNA) & proteins

Ribosomes have 2 subunits

• Small subunit is the part of a ribosome that
mRNA molecules attach to
• Large subunit joins the amino acids together
as tRNA delivers them
Each ribosome has 3 binding sites for
tRNA molecules: the A, P, & E sites
 “Reading” mRNA
tRNA “reads” mRNA in short 3-nucleotide
groups called codons
• Each codon is complementary to the anticodon
on a specific tRNA molecule

To correctly read an mRNA message,

translation must start at the correct location
on the mRNA strand
• This establishes the reading frame, ensuring that
the right groups of 3 letters are “read” as a codon
• The start codon, which marks the beginning of
every protein, is AUG
 Translation: An Overview
Step 1: Initiation Step 2: Elongation Step 3: Termination
• The Start codon is • tRNA “reads” the • The ribosome
located on the mRNA mRNA reaches a Stop codon
• The two ribosomal • A chain of amino • It releases its amino
subunits come acids is created acid chain
together

Initiation Elongation Termination

Translation Animation
 The Genetic Code Table
The Genetic Code table can be used to
determine which amino acid correlates
to each codon in an mRNA molecule

The Genetic Code is degenerate

• This means more than one codon leads to
the same amino acid
• This creates some “wiggle room” for
errors in transcription

BUT – only the last letter in a codon can

be changed without major consequences
• This is called the codon’s wobble base
 Let’s Practice!
Translate the following mRNA molecule into an amino acid
sequence:

5’ A U G C C A U C G C U A U G A 3’
From Plan… … To Protein

A “copy” of the
blueprint is redrawn
with specific The workers use
measurements that that version to
the construction build a new
workers can follow. building.
 Transcription Translation

The nucleic acid it starts

with:

What it ends with:

Where this happens:

Amino acids mRNA Amino acids mRNA

Circle all the things it uses: Ribosomes Nucleotides Ribosomes Nucleotides
tRNA RNA Polymerase tRNA RNA Polymerase
 Consider This…
Every cell in your entire body has the exact
same genome
• This means that all of your cells have the
potential to make ANY of the proteins your
genome encodes!

But…
• Neurons don’t make myosin (a muscle
contraction protein), and…
• Muscle cells don’t make insulin (a protein that
enables glucose entry into your cells), and…
• Pancreatic cells don’t make acetylcholine (a
protein neurons use to communicate with one
another)
 Differential Gene Expression
Differential gene expression is what cells use to selectively express
only the genes that encode proteins they actually need

Differential gene
expression enables
non-specialized stem
cells to differentiate
Stem
into specific types of cell
functional cells.
 Transcription factors are proteins that assist with
differential gene expression.

Repressor proteins block RNA

polymerase, leading to decreased
protein production.

Activator proteins increase the rate of

transcription, leading to increased protein
production.
 Differential Gene Expression & Insulin Resistance
Insulin resistance is a common symptom of
Type 2 diabetes.
• In this condition, pancreatic β cells still make
insulin, but the body’s cells don’t respond to it
• This means the cells are less able to import
glucose

One cause of insulin resistance is a decrease

in the expression of the insulin receptor
gene
• As gene expression decreases, fewer insulin
receptors are made
• The lack of these receptors makes cells unable
to take up glucose
 Mutations
Mutations are changes to the
DNA or mRNA of a gene that is
being used to build a protein
• If mutations are not repaired, the
proteins may be misshapen and
potentially non-functional

Types of mutations
• In a point mutation, a single
nucleotide is incorrect
• In a frameshift mutation, the
reading frame of the mRNA
molecule shifts
 Point mutations occur when an Types of Point Mutations
incorrect nucleotide is used to build a
DNA or mRNA strand. Silent mutation - the changed
nucleotide does NOT impact the
amino acid added to the protein

Missense mutation - the

changed nucleotide leads to the
WRONG amino acid being added
to the protein

Nonsense mutation - the

changed nucleotide leads to the
generation of a STOP codon
 Frameshift mutations are caused by the
insertion / deletion of 1 - 2 nucleotides.

Frameshift mutations
disrupt the ribosome’s
reading frame.
This means the mRNA
nucleotides are “read” in
incorrect groups.

Frameshift mutations can have Frameshift mutations can have

missense effects  the wrong nonsense effects  a new STOP
amino acid is added to the protein. codon to is generated.
 If 3 nucleotides are inserted or deleted at the same time, the
reading frame does not change.

BUT :
The protein that is
generated will still have an
incorrect amino acid
sequence.
“Mutations” in a Sentence:
An Example

Point
mutations

Frameshift
mutations
 What Causes Mutations?
Mutations can arise due to errors in Mutations can arise due to exposure
normal cellular processes. to mutagens in the environment.
 Mutation Detected! If the mutation
cannot be repaired…
Response #1: Response #2:
Attempt DNA
Repair Apoptosis

+
Halt the Cell
Cycle
This prevents mutated
cells from undergoing
mitosis & generating This programmed cell death ensures
mutated offspring. that cells with mutated DNA are
eliminated.
 Diabetes & Mutations
Diabetes has been linked to multiple mutations in in the
insulin-encoding gene (INS gene)

These mutations
affect non-protein-
coding regions of
the gene.

These mutations
affect protein-coding
regions of the gene.
 Mutations in the INS
gene ultimately lead to
changes the structure
of the insulin protein.

What does this picture

show?
The central (colored) circles
represent the normal amino acid
sequence of the insulin protein.
The external circles represent
incorrect amino acids present in
the insulin made by patients with
diabetes.
 To Prepare for Next Class…
 Review your class notes
• Use the eTextbook & Other Helpful Resources to supplement your lecture notes

 Complete the homework assignment and use it to direct your

studying

 Print the slides for Lesson #8: The Cellular Circle of Life