Professional Documents
Culture Documents
1969
Entire genetic code determined
Nucleic Acids
• Nucleic Acids are very long, thread-like polymers, made up of a linear array of monomers
called nucleotides.
• Nucleic acids vary in size in nature
• tRNA molecules contain as few as 80 nucleotides
• Eukaryotic chromosomes contain as many as 100,000,000 nucleotides.
Two types of nucleic acid are found
Deoxyribonucleic acid (DNA)
Ribonucleic acid (RNA)
DNA
deoxyribonucleic acid
nucleic acid that stores genetic information
found in the nucleus of a mammalian cell.
RNA
ribonucleic acid
3 types of RNA in a cell
Ribosomal RNAs (rRNA) are components of ribosomes
Messenger RNAs (mRNA) carry genetic information
Transfer RNAs (tRNA) are adapter molecules in translation
NUCLEOTIDE STRUCTURE
DNA to Protein
DNA acts as a “manager” in the process of making proteins
DNA is the template or starting sequence that is copied into RNA that is then used to make
the protein
Central Dogma
One gene – one protein
This is the same for bacteria to humans
DNA is the genetic instruction or gene
DNA → RNA is called Transcription
RNA chain is called a transcript
RNA → Protein is called Translation
Expression of Genes
Some genes are transcribed in large quantities because we need large amount of this
protein
Some genes are transcribed in small quantities because we need only a small amount of
this protein
Nucleotides as Language
We must start to think of the nucleotides – A, G, C and T as
part of a special language – the language of genes that we
will see translated to
the language of
amino acids in
proteins
Genes as Information
Transfer
A gene is the sequence of nucleotides within a portion of DNA that codes for a peptide or a
functional RNA
Sum of all genes = genome
DNA Replication
Semiconservative
Daughter DNA is a double helix with 1 parent strand and 1 new strand
Found that 1 strand serves as the template for new strand
DNA Template
Each strand of the parent DNA is used as a template to make the new daughter strand
DNA replication makes 2 new complete double helices each with 1 old and 1 new strand
Replication Origin
Site where replication begins
1 in E. coli
1,000s in human
Strands are separated to allow replication machinery
contact with the DNA
Many A-T base pairs because easier to break 2
H-bonds that 3 H-bonds
Note anti-parallel chains
Replication Fork
Bidirectional movement of the DNA replication
machinery
THE REPLICATION FACTORY
DNA replication is an intricate process requiring the concerted action of many different
proteins.
The replication proteins are clustered together in particular locations in the cell and may
therefore be regarded as a small “Replication Factory” that manufactures DNA copies.
The DNA to be copied is fed through the factory, much as a reel of film is fed through a
movie projector.
The incoming DNA double helix is split into two single strands and each original single
strand becomes half of a new DNA double helix. Because each resulting DNA double helix
retains one strand of the original DNA, DNA replication is said to be semi-conservative.
STRAND SEPARATION
To begin the process of DNA replication, the two double helix strands are unwound and
separated from each other by the helicase enzyme.
The point where the DNA is separated into single strands, and where new DNA will be
synthesized, is known as the replication fork.
Single-strand binding proteins, or SSBs, quickly coat the newly exposed single strands.
SSBs maintain the separated strands during DNA replication.
Without the SSBs, the complementary DNA strands could easily snap back together.
SSBs bind loosely to the DNA, and are displaced when the polymerase enzymes begin
synthesizing the new DNA strands.
REPLICATION IN ACTION
The process begins when the helicase enzyme unwinds the double helix to expose two
single DNA strands and create two replication forks.
DNA replication takes place simultaneously at each fork. The mechanism of replication is
identical at each fork.
Proofreading
DNA Repair
For the rare mutations occurring during replication that isn’t caught by DNA polymerase
proofreading
For mutations occurring with daily assault
If no repair
In germ (sex) cells à inherited diseases
In somatic (regular) cells à cancer
Types of Mutation
1. Induced
viruses, UV radiation, some chemicals (nitric acid changes cytosine to uracil)
or mutagens (or carcinogens - benzene, cigarette smoke).
2. Spontaneous
Proofreading mistakes during DNA replication (Base substitutions) - not necessarily
a serious change.
Frame shift mutation (Addition or deletion of a base) - serious change!
A 3 letter code or codon is analogous to three letter words in a sentence.
Original sequence
THE CAT SAW THE DOG
Base or letter substitutions
THE BAT SAW THE DOG
THE CAT SAW THE HOG
THE CAB SAW THE DOG
THE CAT SAW SHE DOG
THE CAT SAD THE DOG
THE CAT SAW THE DOC
Deletions
THE CAT SAW TED OG
THE ATS AWT HED OG
Additions
THE CAT SAW THE ZDO G
THE CMA TAS WTH EDO G
Effect of Mutation
Uncorrected Replication Errors
Mismatch repair
Enzyme complex recognizes mistake and excises newly-synthesized strand and fills
in the correct pairing
Eukaryotes “label” the daughter strand with nicks to recognize the new strand
Separates new from old
Chemical Modifications
Thymine Dimers
Repair Mechanisms
Different enzymes recognize, excise different mistakes
DNA polymerase synthesizes proper strand
DNA ligase joins new fragment with the polymer
STEP 2 - TRANSCRIPTION
Transcription
The region of the double-stranded DNA corresponding to a
specific gene is copied into an RNA molecule, called
messenger RNA (mRNA).
RNA differs from DNA
Ribose is the sugar rather than deoxyribose – ribonucleotides
U instead of T; A, G and C the same
Single stranded
Can fold into a variety of shapes that allows RNA to have structural and
catalytic functions
RNA Differences
Transcription
Similarities to DNA replication
Open and unwind a portion of the DNA
1 strand of the DNA acts as a template
Complementary base-pairing with DNA
Differences
RNA strand does not stay paired with DNA
DNA re-coils and RNA is single stranded
RNA is shorter than DNA
RNA is several 1000 bp or shorter whereas DNA is 250 million bp long
RNA Polymerase
Catalyzes the formation of the phosphodiester bonds between the nucleotides (sugar to
phosphate)
Uncoils the DNA, adds the nucleotide one at a time in the 5’ to 3’ fashion
Uses the energy trapped in the nucleotides themselves to form the new bonds
Template to Transcripts
The RNA transcript is identical to the NON-template strand
with the exception of the T’s becoming U’s
RNA Elongation
Reads template 3’ to 5’
Adds nucleotides 5’ to 3’ (5’ phosphate to 3’ hydroxyl)
Synthesis is the same as the leading strand of DNA
Differences in DNA and RNA Polymerases
RNA polymerase adds ribonucleotides not deoxynucleotides
RNA polymerase does not have the ability to proofread what they transcribe
RNA polymerase can work without a primer
RNA will have an error 1 in every 10,000 nucleotides (DNA is 1 in 100,000,000 nucleotides)
Types of RNA
messenger RNA (mRNA) – codes for proteins
ribosomal RNA (rRNA) – forms the core of the ribosomes, machinery for making proteins
transfer RNA (tRNA) – matches code for amino acid on mRNA and positions the right amino
acid in place during protein synthesis
STEP 3 – TRANSLATION
RNA to Protein
Translation is the process of turning mRNA into protein
Translate from one “language” (mRNA nucleotides) to a second “language” (amino acids)
Genetic code – nucleotide sequence that is translated to amino acids of the protein
DNA Code
Reading Frames
Translation can occur in 1 of 3 possible reading frames, dependent on where decoding
starts in the mRNA
Attachment of AA to tRNA
Aminoacyl-tRNA synthase is the enzyme responsible for linking the amino
acid to the tRNA
A specific enzyme for each amino acid and not for the tRNA
Ribosomes
Complex machinery that controls protein synthesis
2 subunits
1 large – catalyzes the peptide bond formation
1 small – binds mRNA and tRNA
Contains protein and RNA
rRNA central to the catalytic activity
Folded structure is highly conserved
Protein has less homology and may not be as important
Ribosome Structures
Ribosomal Subunits
Ribosomal Movement
4 binding sites
mRNA binding site
Peptidyl-tRNA binding site (P-site)
Holds tRNA attached to growing end of the peptide
Aminoacyl-tRNA binding site (A-site)
Holds the incoming AA
Exit site (E-site)
Summary