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Molecular Biology
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Chapter 4
Transcription

Written by: Radia AITOUNY

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Table of Contents:

I. Transcription key players

II. Transcription: Process
1. Initiation
2. Elongation
3. Termination
III. Post mRNA synthesis: mRNA maturation processes

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Introduction

• Protein synthesis: proteins make up all living materials.

• Proteins are composed of amino acids: there are 20 different
amino acids.
• Different proteins are made by combining these 20 amino acids in
different combinations.

• Transcription: process by which the information in a strand of DNA is

copied into a new molecule of messenger RNA (mRNA)
• Less than 2% of the human genome can be transcribed into mRNA. A messenger
RNA (mRNA) molecule is synthesized from the DNA and directs the formation of
the protein.

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I. Transcription Key Players

1. Ribosomes
2. Amino Acid
3. tRNA
4. Gene
5. RNA Polymerase
6. mRNA

1. Ribosomes

• The ribosome is a complex cellular machine. It is largely made up of

ribosomal RNA (rRNA) and proteins.

• Ribosomes consist of two major components: the small (30S) and the large
(50S) subunits. The two are bound to each other.
• Each ribosomal subunit has 3 binding sites for tRNA:
- A (aminoacyl) site, which accepts the incoming aminoacylated tRNA
- P (peptidyl) site, which holds the tRNA with the nascent peptide chain
- E (exit) site, which holds the deacylated tRNA before it leaves the
ribosome

2. Amino Acid

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• Amino Acids are the precursors to proteins.

• Amino Acids are organic compounds that contain:
- An amine group (NH2)
- A carboxylic acid group (COOH)
- A variable side chain (R)

• 20 amino acids make proteins found in the human body. Humans can produce 10
AA. The rest come from the food we eat.

3. tRNA: transfer RNA

• tRNA is a small RNA molecule that participates in protein synthesis. It has about
76 to 90 nucleotides.
• tRNA serves as the physical link between mRNA and the amino acid.

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• Each tRNA has 3 nucleotides (ex: AUG) that are complementary to the codon in
mRNA, and codes for a specific amino acid.

4. Gene

• A gene is the basic physical and functional unit of heredity. A gene’s DNA
is transcribed into mRNA which is later translated into a protein.
• The human genome contains an estimated 20,000 to 25,000 genes.
However, only about 1% of the genome is made up of protein-coding
genes. The rest is made up of non-coding DNA, and has other important
functions such as regulating the transcription and translation

• Every gene consists of two functional components functional units:

- The promoter region
- The coding region
• The promoter region is the 5’ region of the DNA, upstream of the coding
sequence.
- Cis-acting elements are DNA sequences in the vicinity of the structural
portion of a gene, which are required for gene expression. Trans-acting
factors are usually protein factors that bind to the cis-acting sequences to
control gene expression

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• The coding region has exons and introns:

- Exons: coding sections of the DNA that are translated into protein
- Introns: sections of DNA that do not code for proteins
• Gene enhancers/silencers
- Enhancers: short DNA sequence (50-1500 base pairs) capable of binding
transcription regulation factors (called activators). They increase the
transcription of a particular gene.
- Silencers: short DNA sequence capable of binding transcription regulation
factors (called repressors). They decrease or inhibit transcription.
• Gene insulators
- Insulators are thought to act as barriers, preventing enhancers and
silencers from regulating neighboring genes.

5. RNA Polymerase
• RNA polymerase is the enzyme that produces the RNA molecules. It
produces at least 5 types of RNA for 2 functions
- Production of proteins: mRNA, tRNA, rRNA
- Regulation: miRNA (microRNA), catalytic RNA
• There are at least 3 types of RNA polymerase:
- RNA polymerase I: transcribes ribosomal RNA (rRNA)
- RNA polymerase II: transcribes DNA into pre-messenger mRNA
- RNA polymerase III: transcribes short RNA’s (such as tRNA)
• RNA polymerase moves in 3’ à 5’ direction, and synthesizes the new
mRNA strand in 5’ à 3’ direction.

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6. mRNA: messenger RNA

• mRNA is a single-stranded molecule that is complementary to one of the
DNA strands of a gene.
• The mRNA is an RNA version of the gene (DNA).
• In eukaryotes, protein translation can only start at Methionine codon: AUG.

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II. Transcription: process

1. Initiation
• In the nucleus, RNA polymerase recognizes the recognition sites causing it to
bind to the promoter (the start of a gene).
• The RNA Polymerase then separates the DNA into single strands so the template
strand can be read in the 3’ à 5’ direction.
• This is done by breaking the hydrogen bonds between complementary DNA
nucleotides.

2. Elongation
• Pre-mRNA nucleotides are quickly paired with their complementary bases which
correspond with the template strand of DNA.
• The RNA polymerase reads the DNA template in a 3’ à 5’ direction, while the
synthesis of pre-mRNA moves in the 5’ à 3’ direction.
• Only one of the two strands of DNA is transcribed: it’s called the template.
• The mRNA product is complementary to the template strand. Therefore, it is
almost identical to the non-transcribed strand (coding strand): the only difference
is that the mRNA contains Uracil (U) instead of Thymine (T) found in DNA.

3. Termination
When the RNA polymerase reaches the terminator, it signals the RNA
polymerase to stop and release from the DNA. Once separated the two DNA
strands come back together and reform the double helix. The newly formed pre-
mRNA molecule is then released.

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III. Post mRNA synthesis: mRNA maturation processes

Three main processing steps:
- Capping at the 5’ end
- Addition of a polyA tail at the 3’ end
- Splicing to remove introns

1. The CAP
Shortly after RNA Polymerase II initiates transcription the 5’ end of the nascent
RNA is capped with 7-methylguanylate (the guanosine is methylated on the 7th
carbon).

2. Polyadenylation (polyA tail)

The polyA tail is a stretch of RNA that only has adenine bases. This tail protects
the mRNA from the degradatory action of nucleases. (250 A long in mammals)

3. Splicing
The last step of mRNA maturation consists of removing introns and joining exons.
The spliceosome (large ribonucleoprotein complex) removes introns from
transcribed pre-mRNA.

Alternative splicing is a process that enables a messenger RNA (mRNA) to direct

synthesis of different protein variants (isoforms). In humans up to 10 different
proteins can be traced to a single gene; 20,000-25,000 genes produce between
80,000 and 400,000 proteins.

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