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Nucleic Acids and
Protein Synthesis
THE FUNCTIONS OF DNA
DNA has three roles/purposes/functions: 1. Storing information – genes are
segments of DNA that carry messages to make proteins.
2. Copying information – so that when cells divide, all cells get a complete copy of the genetic material.
3. Transmitting information – DNA is passed from parents to offspring.
THE STRUCTURE OF DNA
Nucleotides are the building blocks of DNA.
Each nucleotide contains a phosphate group, a five carbon sugar, and a nitrogen base. ◦ The five carbon sugar is
called deoxyribose. ◦ Covalent bonds hold the
sugar of one nucleotide to the phosphate group of another nucleotide to form chains.
THE STRUCTURE OF DNA
The full name of DNA is deoxyribonucleic acid.
Every nucleotide has the same sugar molecule and phosphate group, but each nucleotide contains one of four nitrogen bases. ◦ The four nitrogen bases that
make up DNA are adenine, thymine, guanine, and cytosine.
THE STRUCTURE OF DNA
Adenine and Guanine are called
purines. Purines have 2 rings of
carbon and nitrogen atoms.
Thymine and Cytosine are called
pyrimidines. Pyrimidines have a
single ring of carbon and nitrogen
atoms.
CHARGAFF’S RULE
1949-Erwin Chargaff showed that in DNA, the number of adenines equal the number of thymines, AND the number of cytosines equal the number of guanines. However, the amount of each nucleotide was not the same among different organisms.
Base-pairing / Chargaff’s rule:
◦ Adenine will always pair with Thymine; A and T are complimentary.
◦ Cytosine will always pair with Guanine; C and G are complimentary.
THE DNA MOLECULE IS A
DOUBLE HELIX
Rosalind Franklin and Maurice Wilkins used X
ray diffraction to take first picture of DNA.
Determined a two dimensional picture of DNA’s
structure.
James Watson and Francis Crick – 3-D shape
of DNA being 2 strands of nucleotides that form
a spiral staircase or double helix.
THE DNA MOLECULE IS A
DOUBLE HELIX
DNA is a twisted ladder with alternating patterns of phosphates and sugars making the sides of the ladder. ◦ Each rung is a purine/pyrimidine pair held together by hydrogen bonds.
THE DNA MOLECULE IS A
DOUBLE HELIX The base pair rules tell us what the rungs
can be, A and T or G and C.
Each strand of the double helix is complementary to each other; the sequence of 1 strand determines the sequence of the other.
The two strands of DNA in the double-helix are antiparallel – they run in opposite directions.
HOW DNA IS COPIED
DNA is double stranded – base pairing allows for easy copying; one strand serves as a template for a new strand.
DNA replication – the process of making a new DNA strand. ◦ Occurs before cells divide.
◦ Ensures that when cells divide, each cell produced has an entire copy of the organism’s DNA.
DNA double helix is unzipped by an enzyme called a helicase. Helicase breaks hydrogen bonds linking the nitrogen bases. ◦ Occurs at the replication forks of the double helix.
At the replication fork; an enzyme called DNA
polymerase moves along the strands, reading
the nitrogen base of each nucleotide, and
adding the complementary nucleotide to the
new strand. ◦ Remember that A and T are complimentary; C and G
are also complimentary.
HOW DNA IS COPIED
Telomeres – tips of chromosomes where DNA is hard to replicate. ◦ Telomerase – enzyme that adds repeated DNA
sequences to the ends of the chromosomes to prevent loss of the telomere DNA during DNA replication.
HOW DNA IS COPIED
Replication in prokaryotes– single circular chromosome; replication begins and proceeds from ONE location on the chromosome.
Replication in eukaryotes – many linear chromosomes; replication begins and proceeds from hundreds of locations on each chromosome.
HOW DNA IS COPIED
Gene – segment of DNA that codes for a protein.
Cells transfer the information found within the genes on DNA into a set of working instructions for use in building proteins.
This working set of instructions of the gene is called ribonucleic acid or RNA. ◦ RNA is a nucleic acid made of chains of nucleotides, just like DNA.
THE PATH OF GENETIC
INFORMATION
THE PATH OF GENETIC
INFORMATION
RNA is a single strand of nucleotides; DNA is
double stranded.
The sugar in RNA is a 5 Carbon sugar called
ribose; DNA’s sugar is deoxyribose.
RNA does not contain Thymine, but has replaced
Thymine with the base Uracil.
DNA compared to RNA
DNA RNA
How many strands?
2 1
Nucleotide subunit
Deoxyribose sugar
Ribose sugar
Bases Thymine (T) Adenine (A) Guanine (G) Cytosine (C)
Uracil (U) Adenine (A) Guanine (G) Cytosine (C)
Phos-
phate Group
Deoxy-ribose Sugar
Nitro-gen Base Phos-
phate Group
Ribose Sugar
Nitro-gen Base
T – A G – C
U – A G – C
THE PATH OF GENETIC
INFORMATION
Three forms of RNA are messenger RNA (mRNA),
transfer RNA (tRNA), and ribosomal RNA (rRNA).
All 3 RNA’s are responsible for processing the
information in a gene into protein. The process of
transferring the information in genes to proteins is
called gene expression.
THREE TYPES OF RNA
mRNA – used as a blueprint or template for a
protein; carries DNA’s information from the nucleus
to site of translation (ribosomes in cytoplasm).
tRNA – decodes mRNA into amino acid
sequences.
rRNA – RNA part of a ribosome’s structure (the
other component of ribosomes is protein).
Gene expression occurs in 2 stages. ◦ The first, transcription is where DNA is transferred to
mRNA.
◦ The second stage; translation, is when the information in
mRNA is used to make protein.
THE PATH OF GENETIC
INFORMATION
TRANSCRIPTION: MAKING RNA
Transcription takes place inside the nucleus.
Transcription begins when RNA polymerase binds to the beginning of a gene on a region of DNA.
TRANSCRIPTION: MAKING RNA
The region of DNA to which RNA polymerase
binds is called a promoter. Promoters are
sequences of DNA that act as a start signal.
The RNA polymerase begins to unzip and
separate the double helix.
The polymerase uses only one of the DNA
strands as a template for mRNA – The non-coding
or complimentary strand is the template for
mRNA synthesis.
TRANSCRIPTION: MAKING RNA
Follows the same base pairing rules as replication
except Uracil is used in place of Thymine.
RNA nucleotides are added one at a time in the
active site of the RNA polymerase.
DNA reforms the double helix following the RNA
Polymerase.
Transcription occurs at about 60 nucleotides per
second.
Terminator - the stop signal in the sequence of
DNA – RNA Polymerase detaches here.
TRANSCRIPTION: MAKING RNA
Coding DNA: CTC TTG ATC ATG
Non-coding/complimentary DNA:
GAG AAC TAG TAC
RNA: CUC UUG AUC AUG
http://207.207.4.198/pub/flash/26/transmenu_s.swf
Post –transcriptional mRNA processing
RNA editing - mRNA must be processed, or
prepared for the next phase of gene expression,
translation. ◦ Introns – non-coding gene regions that are cut out of the
RNA molecule in the nucleus.
◦ Exons – expressed sequences of the RNA molecule;
codes for a protein.
◦ The introns are cut out and the exons are spliced together to make a final mRNA.
Post –transcriptional mRNA processing
A protective cap and tail are then added before the mRNA leaves the nucleus through one of the pores and heads to the cytoplasm where translation will occur on ribosomes.
THE GENETIC CODE
Instructions on mRNA are written as a series of
three nucleotide sequences called a codon.
Each codon (set of three nucleotides) corresponds
to a certain amino acid or a stop signal. ◦ 64 possible codon combinations.
◦ Genetic code – collection of codons of mRNA, each of
which directs the incorporation of a particular amino acid
during protein synthesis.
Codon Table
START AND STOP CODONS
Start codon or initiator codon – AUG; cues the
start of translation by inserting a methionine. ◦ Thus, all proteins begin with the amino acid methionine.
Translation proceeds until a stop codon is
reached. ◦ Stop codon – triggers the end of translation.
TRANSLATION: MAKING PROTEINS
Translation – when ribosomes in the cytoplasm
use the sequence of codons in mRNA to assemble
amino acids into polypeptide chains.
tRNA is a single stranded RNA folded into a
compact shape with three loops. ◦ One loop has a three nucleotide sequence (called an
anticodon) that is complementary to one of the 64
codons.
◦ Each tRNA carries one amino acid.
TRANSLATION: MAKING PROTEINS
TRANSLATION: MAKING PROTEINS
tRNA bonds with mRNA at the codon/anticodon
site by hydrogen bonds. ◦ Every tRNA carries a particular amino acid that
corresponds to the particular codon.
Once the amino acid has been added to the
growing polypeptide chain, the tRNA is released.
Many amino acids link to form peptides – once a
peptide is folded into its proper shape it is
considered a protein.
Central Dogma
Central dogma of molecular biology –
information is transferred from DNA to RNA to
protein. ◦ This allows gene expression, or DNA, RNA, and proteins
working together to put the genetic information contained
in cells into action.
Regulation of gene expression -
Prokaryotes
Prokaryotic gene expression is regulated by DNA
binding proteins. ◦ These regulatory proteins help switch genes on and off.
◦ Operon – group of prokaryotic genes regulated together.
Promoter – DNA sequence where RNA polymerase binds to
begin transcription.
Operator – DNA sequence where regulatory proteins can
bind to repress transcription.
Regulation of gene expression -
Eukaryotes
Eukaryotic gene expression is also regulated by
DNA binding proteins, however, eukaryotes
typically regulate individual genes, not groups of
them. ◦ TATA box – short sequence of DNA that marks the
beginning of a gene; used to help position RNA
polymerase.
Regulation of gene expression -
Eukaryotes
Transcription factors – proteins
that help regulate gene
expression by binding DNA
promoter/enhancer sequences
and blocking or activating
transcription.
◦ Promoter/enhancer –
sequences of DNA with binding
sites for multiple transcription
factors.
Regulation of gene expression -
Eukaryotes Not all genes are expressed in all eukaryotic cells.
◦ Cell specialization – all cells in multicellular organisms
contain all of the organism’s DNA, yet they only
transcribe and translate part of it.
Ex. Liver cells only transcribe and translate liver specific
genes while skin cells only transcribe and translate skin
specific genes.
◦ RNA interference (RNAi) – used to regulate gene
expression in eukaryotes.
During RNAi, microRNAs (miRNAs) bind to transcribed
mRNA to block it from being translated into protein.
Regulation of gene expression -
Eukaryotes
Differentiation – when gene regulation allows eukaryotic cells to become specialized in structure and function. ◦ Occurs during embryonic development.
◦ Homeotic genes aka master control genes – specific group of genes that controls the identity of body parts in embryos. Homeobox genes – code for transcription factors that
activate other genes important for development and differentiation. ◦ In flies, these are called Hox genes.
The environment also plays a role in the regulation of prokaryotic and eukaryotic gene expression.