Gene Expression: The Flow of

Information from DNA

to RNA to Protein
BY
YEBOAH KWAKU OPOKU Ph.D.
DEPARTMENT OF BIOLOGY EDUCATION
FACULTY OF SCIENCE EDUCATION
UNIVERSITY OF EDUCATION, WINNEBA
Outline

• The Genetic Code

• Transcription: From DNA to RNA
• Translation: From mRNA to Protein
 Introduction
• The term gene expression refers to the process by which
information contained in genes is decoded to produce other molecules
that determine the phenotypic traits of organisms.
• The process is initiated when the information contained in the base
sequence of DNA is copied into a molecule of RNA, and the process
culminates when the molecule of RNA is used to determine the linear
order of amino acids in a polypeptide chain.
• Therefore genetic information flows from DNA to RNA to protein.
• This statement was set forward as the “ Central Dogma” of
molecular biology by Francis Crick in 1957
• The Central Dogma maintains that genetic information
flows in two distinct stages
• The conversion of DNA-encoded information to its
RNA-encoded equivalent is known as transcription.
• In the second stage of gene expression, the cellular
machinery translates mRNA into its polypeptide
equivalent in the language of amino acids.
• This decoding of nucleotide information to a sequence
of amino acids is known as translation
• The protein made is called the gene product.
• Translation takes place on molecular workbenches
called ribosomes
• The ribosomes are composed of proteins and ribosomal
RNAs (rRNAs), and it depends on the “dictionary”
known as the genetic code
• The genetic code defines each amino acid in terms of
specific sequences of three nucleotides.
• Translation also depends on transfer RNAs (tRNAs) -
place specific amino acids at the correct position in a
growing polypeptide chain
 The Genetic Code
• The language of nucleic acids is written in four nucleotides—A, G, C,
and T in the DNA and A, G, C, U in the RNA dialect
• However, the language of proteins is written in amino acids
• How can four nucleotides encode 20 amino acids?
• If only one nucleotide represented an amino acid, there would be only
four amino acids
• If two nucleotides represented each amino acid, there would be
42=16 possible combinations of couplets.
• Groups of three nucleotides in a row would provide 43=64 different
triplet combinations, more than enough to code for all the amino acids
 Colinearity of genes and proteins
• Most genes contain the information for the synthesis of only
one polypeptide chain.

• Furthermore, the linear order of nucleotides in a gene

determines the linear order of amino acids in a polypeptide

• This point was first proved by studies of the tryptophan

synthase gene trpA in E. coli, a gene in which many mutations
had been obtained and accurately mapped genetically by
Yanofsky
• More important, the order of the mutations in the
genetic map was the same as the order of the affected
amino acids in the polypeptide chain

• This attribute of genes and polypeptides is called

colinearity, which means that the sequence of base
pairs (bp) in DNA determines the sequence of amino
acids in the polypeptide in a point-to-point, manner
Colinearity of genes and proteins
 Evidence for a triplet
code by Crick and Brenner

• The restoration of gene function by

one mutation canceling another in
the same gene is known as
intragenic suppression
• Crick and Brenner assumed that
each codon is a trio of nucleotides
and that for each gene there is a
single starting point.
• This starting point establishes a
reading frame
• Each nucleotide triplet is
called a codon.
• Each codon, designated by
the bases defining its three
nucleotides, specifies one
amino acid.
• For example, GAA is a codon
for glutamic acid (Glu), and
GUU is a codon for valine
(Val)
• The three stop codons that
terminate translation are
known as nonsense codons.
• The codons are nonoverlapping- e.g 5’ GAAGUUGAA 3’,
the first three nucleotides (GAA) form one codon; (GUU)
form the second; each nucleotide is part of only one codon.
• The code is degenerate, meaning that more than one codon
may specify the same amino acid.
• The cellular machinery scans mRNA from a fixed starting
point that establishes a reading frame.
• The nucleotide triplet AUG, which specifies the amino acid
methionine, serves in certain contexts as the initiation codon,
marking where in an mRNA the code for a particular
polypeptide begins.
 Transcription: From DNA to RNA
• Transcription is the process by which the polymerization of
ribonucleotides guided by complementary base pairing produces an
RNA transcript of a gene.
• The template for the RNA transcript is one strand of that portion of
the DNA double helix that composes the gene.
• Transcription can be divided into 3 successive phases of initiation,
elongation, and termination.
• The enzyme RNA polymerase catalyzes transcription.
• DNA sequences near the beginning of genes, called promoters, signal
RNA polymerase where to begin
• Sequences in the RNA products, known as terminators, tell RNA
polymerase where to stop transcription.
RNA polymerase recognizes and binds to promoters on the double-stranded DNA at the beginning of
the gene
RNA polymerase unwinds part of the double helix, exposing unpaired bases on the template strand.
RNA polymerase aligns the first two ribonucleotides of the new RNA, which will be at the 5’ end of
the final RNA product.
RNA polymerase catalyzes the formation of a phosphodiester bond between the first two
ribonucleotides.
The RNA polymerase releases the δ subunit marking the end of initiation
 The core enzyme now moves along the chromosome, unwinding the double helix to
expose the next single stranded region of the template.
 The enzyme extends the RNA by linking a ribonucleotide positioned by
complementarity with the template strand to the 3’ end of the growing chain
 RNA sequences that signal the end of transcription are known as terminators.
 There are two types of terminators: intrinsic terminators, which cause the RNA polymerase
core enzyme to terminate transcription on its own, and extrinsic terminators, which require
proteins other than RNA polymerase such as polypeptide known as rho to bring about
termination.
 All terminators are sequences in the mRNA that are transcribed from specific DNA regions.
 Terminators often form hairpin loops in which nucleotides within the mRNA pair with
nearby complementary nucleotides.
 Upon termination, RNA polymerase and a completed RNA chain are released from the DNA.
 The product of transcription is a Single-Stranded Primary Transcript
 The bases in the primary transcript are complementary to the bases between the
initiation and termination sites in the template strand of the gene.
 The ribonucleotides in the primary transcript include a start codon, the codons
that specify the remaining amino acids of the polypeptide, and a stop codon
 RNA processing after transcription produces
a mature mRNA in eukaryotes
• Some RNA processing in eukaryotes modifies only the 5’ or 3’ ends
of the primary transcript, leaving the information content of the rest of
the mRNA untouched.
• Other processing deletes blocks of information from the middle of
the primary transcript, so the content of the mature mRNA is
related, but not identical, to the complete set of DNA nucleotide pairs
in the original gene
• Firstly, the nucleotide at the 5’ end of a eukaryotic mRNA is G in
reverse orientation from the rest of the molecule connected
through a triphosphate linkage to the first nucleotide in the primary
transcript.
• This “backward G” is not transcribed from the DNA but
added by a special capping enzyme to the primary transcript
after polymerization of the transcript’s first few nucleotides.

• Enzymes known as methyl transferases then add methyl (–

CH 3 ) groups to the backward G and to one or more of the
succeeding nucleotides in the RNA, forming a so-called
methylated cap
• Secondly, in a large majority of eukaryotic mRNAs, the
3’ end consists of 100–200 A’s, referred to as a poly-A
tail
• A ribonuclease cleaves the primary transcript to form a
new 3’ end; cleavage depends on the sequence
AAUAAA, which is found in poly-A-containing
mRNAs 11–30 nucleotides upstream of the position
where the tail is added.
• Next the enzyme poly-A polymerase adds A’s onto the
3’ end exposed by cleavage.
• It was found that the DNA nucleotide sequences of many
eukaryotic genes are much longer than their corresponding
mRNAs
• Suggesting that RNA transcripts, in addition to receiving a
methylated cap and a poly-A tail, undergo extensive
internal process
• Sequences found in both a gene’s DNA and the mature
messenger RNA are called exons (“expressed regions”).
• The sequences found in the DNA of the gene but not in the
mature mRNA are known as introns
The process of removing introns or noncoding part of a gene
from an RNA transcript is called RNA splicing
 Translation: From mRNA to Protein
• Translation is the process by which the sequence of
nucleotides in a messenger RNA directs the assembly of the
correct sequence of amino acids in the corresponding
polypeptide.
• Translation takes place on ribosomes that coordinate the
movements of transfer RNAs carrying specific amino acids
with the genetic instructions of an mRNA.
• Transfer RNAs (tRNAs) serve as adaptor molecules that
mediate the transfer of information from nucleic acid to protein
 The structure of tRNA
• Transfer RNAs are short, single-stranded RNA molecules 74–
95 nucleotides in length
• Several of the nucleotides in tRNAs contain modified bases
produced by chemical alterations of the principal A, G, C,
and U nucleotides
• Each tRNA carries one particular amino acid, and cells
must have at least one tRNA for each of the 20 amino acids
specified by the genetic code
• At the other end of the L, where the
5’ and 3’ ends of the tRNA strand are
found, enzymes known as
aminoacyltRNA synthetases connect
the tRNA to the amino acid that
corresponds to the anticodon
The translation process
Translation consists of three phases:
Initiation phase that sets the stage for polypeptide
synthesis
Elongation, during which amino acids are added to a
growing polypeptide
Termination phase that brings polypeptide synthesis to a
halt
 In eukaryotes, the larger polypeptide made before it is
cleaved into smaller polypeptides is often called a
polyprotein
 The addition of chemical constituents, such as phosphate
groups, methyl groups, or even carbohydrates, to specific
amino acids may also modify a polypeptide after translation
 Such cleavages and additions are known as
posttranslational modifications.
THE END