DNA Replication

• Semiconservative – each strand in the double helix acts as a

template for synthesis of a new, complementary strand
• DNA polymerase – enzymes involved in synthesis of new
DNA, synthesizes DNA is the 5’ to 3’ direction
• During DNA replication, one new strand (the leading strand) is
made as a continuous piece. The other (the lagging strand) is
made in small pieces.
• DNA replication requires other enzymes in addition to DNA
polymerase, including DNA primase, DNA helicase, DNA
ligase, and topoisomerase.
Transcription:
DNA to mRNA
Transcription
• First step in gene expression
• DNA acts as a template
• Copies gene’s DNA sequence to make an RNA molecule
• RNA polymerases – enzymes which link nucleotides to
form an RNA strand
• Three stages: initiation, elongation, termination
DNA vs. RNA
• Ribose = 2’ is hydroxylated
DNA vs. RNA
• Thymine = demethylated, Uracil
DNA vs. RNA
• RNA looks like a single strand of DNA but uses ribose in place
of deoxyribose, and uracil in place of thymine
RNA Polymerase II
• Functions like that of DNA polymerase (difference: nucleotides
being used)
• Uses a single strand of DNA as template
• Can only add RNA nucleotides (A, U, G, C)

• Creates a complementary
strand of RNA
= mRNA transcript
Stages of Transcription:
Initiation
Elongation
Termination
Stages of Transcription:
Initiation
• RNA polymerase binds to a sequence of DNA called the
promoter (TATA box)
• Separates DNA strands, to obtain the single-strand
template needed for transcription
Stages of Transcription:
Elongation
• RNA polymerase builds an RNA molecule out of
complementary nucleotides
• The chain grows from 5’ to 3’
• This RNA transcript carries the same information as the
non-template strand DNA however, it contains the base
uracil (U) instead of thymine (T)
Stages of Transcription:
Termination
• Terminators – sequences along the DNA strand, signals that
RNA is complete
• Causes the transcript to be released from the RNA
polymerase
mRNA processing:
Splicing
• Removal of introns from the mRNA transcript
• Spliceosome – locates introns, cuts them, fuses the
remaining parts of the mRNA
mRNA processing:
Protection
• Nucleotide sequences are added to each end of the
mRNA
• Protection from degradation that can occur outside of the
nucleus (i.e. enzymes, exonucleases)
• 5’ cap = G nucleotide
• 3’ end = long sequence of A nucleotides (poly-A tail)
Transcription happens for individual
genes. . .
• Cells carefully regulate transcription
• Not all genes are transcribed all the time!
• Transcription is controlled individually for each
gene
Application:
RNA Interference
• Medical therapy used to prevent a number of
diseases
• Disrupt the production of harmful proteins by
intercepting and incapacitating mRNA
transcripts before they go to the ribosomes
CCT CTT TGC ACT CGG ATC GTA CGC DNA strand

Replication

Complementary
DNA strand

Transcription

mRNA
CCT CTT TGC ACT CGG ATC GTA CGC DNA strand

Replication

Complementary
GGA GAA ACG TGA GCC TAG CAT GCG DNA strand

Transcription

GGA GAA ACG UGA GCC UAG CAU GCG mRNA

…what happens to the RNA transcript?
 Translation:
mRNA to Protein
Translation
• Protein synthesis
• RNAs : mRNA, tRNA, rRNA
• mRNA – carries the coded instructions for protein synthesis
• tRNA – brings amino acids to the ribosome so it can build
proteins
• rRNA – makes up ribosomes;m P site and A site
• The Genetic Code = codons; series of three nucleotides
on the mRNA*