chapter 3 dna & rna protein synthesis lecture 6 marieb’s human anatomy and physiology marieb ...
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Chapter 3DNA & RNA
Protein SynthesisLecture 6
Marieb’s HumanAnatomy and
Physiology
Marieb Hoehn
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Lecture Overview
• The Genetic Information
• Structure of DNA/RNA
• DNA Replication
• Overview of protein synthesis
• Transcription
• Translation
• The genetic code
• The fate of cellular proteins
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Overview of the Genetic Information
Genetic information of the cell is found in the nucleus within chromosomes.
Chromosomes contain DNA and protein (chromatin).
Several orders of packing are required to fit the 2 meters of DNA inside!
DNA must be unpacked before its instructions can make cellular components.
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Packing of the Genetic Information
Figure from: Martini, “Human Anatomy & Physiology”, Prentice Hall, 2001
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
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Some Definitions…
Gene – segment of DNA that codes for a protein or RNA- About 30,000 protein-encoding genes in humans- DNA’s instructions are ultimately responsible for the ability of the cell to make ALL its components
Genetic information – instructs cells how to construct proteins; stored in DNA
Genome – complete set of genes of an organism- Human Genome Project was completed in 2001- Genomes of other organisms are important also
Genetic Code – method used to translate a sequence of nucleotides of DNA into a sequence of amino acids
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Structure of Nucleic Acids
Recall that a nucleotide is made up of three components:
- A five-carbon sugar - A nitrogenous base - A phosphate group
What sugar is this?
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
1
34
5
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Structure of Nucleic Acids
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
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Structure of Nucleic Acids
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
Purines: Adenine and Guanine (double ring)
Pyrimidines: Cytosine, Thymine, and Uracil (single ring)
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Structure of Nucleic Acids
5'
3'
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
The linear sequence of nucleotides in a nucleic acid chain is commonly abbreviated by a one-letter code, e.g. A-T-G-C-C, with the 5’ end of the chain at the left; we always read 5’ 3’
A
T
A
T
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Structure of DNA
A single strand of DNA is a polynucleotide chain connected by a sugar-phosphate backbone.
BUT…a complete molecule of DNA is a DOUBLE-STRANDED HELIX.
How is this accomplished?
Notice that the DNA strand has an ‘orientation’, i.e. 5' 3 '
5'
3'
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Structure of DNA
A double-stranded DNA molecule is created by BASE-PAIRING of the nitrogenous bases via HYDROGEN bonds.
Notice the orientation of the sugars on each stand.
*DNA is an antiparallel, double-stranded polynucleotide helix
5'3'
5' 3'
Figure from: Hole’s Human A&P, 12th edition, 2010
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Structure of DNA
Base pairing in DNA is VERY specific. - Adenine only pairs with Thymine (A-T) - Guanine only pairs with Cytosine (G-C)
Note that there are:
- THREE hydrogen bonds in G-C pairs
- TWO hydrogen bonds in A-T pairs
- A purine (two rings)base hydrogen bonds with a pyrimidine base (one ring)
Figure from: Martini, “Human Anatomy & Physiology”, Prentice Hall, 2001
Complementary base pairing…
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Structure of DNA
Hydrogen bonding between bases of DNA
Figures from: Alberts et al., Essential Cell Biology, Garland Press, 1998
More electronegative
H ON
( - )
( + )( - )
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Structure of DNA
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
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Structure of Genetic Information - Review
• two antiparallel polynucleotide chains
• hydrogen bonds hold nitrogenous bases together
• bases pair specifically (A-T and C-G)
• forms a helix
• DNA wrapped about histone proteins forms chromosomes
Figure from: Hole’s Human A&P, 12th edition, 2010
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DNA ReplicationThe precise, accurate replication of DNA is ESSENTIAL to cellular health and viability.
DNA replication occurs during INTERPHASE of the cell cycle (in S phase).
Figure from: Martini, “Human Anatomy & Physiology”, Prentice Hall, 2001
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DNA Replication
Figure from: Martini, “Human Anatomy & Physiology”, Prentice Hall, 2001
THINGS TO NOTE:
1. Replication fork is asymmetrical
2. New strands are synthesized in a 5’ to 3’ direction
3. DNA polymerase has a proofreading function (1 mistake in 109 nucleotides copied!)
4. Semi-conservative replication
5’
5’
5’
5’
3’
3’
5’
3’
3’
3’
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RNA (Ribonucleic Acid)
RNA, like DNA, is a polynucleotide with a sugar, a phosphate, and a nitrogenous base.
However, RNA has some very important differences:
- uses the pentose sugar, ribose
- uses the nitrogenous base, uracil (U) , in place of thymine (T)
- usually exists as a single-stranded molecule
What base do you think Uracil is capable of hydrogen bonding with?
Figure from: Hole’s Human A&P, 12th edition, 2010
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mRNA Molecules
Messenger RNA (mRNA) -• delivers copy of genetic information from nucleus to the cytoplasm
• single polynucleotide chain
• formed beside a strand of DNA
• RNA nucleotides are complementary to DNA nucleotides (but remember, no thymine in RNA; replaced with uracil)
• making of mRNA is transcription
Figure from: Hole’s Human A&P, 12th edition, 2010
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tRNA Molecules
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
Transfer RNA (tRNA) – the adapters in translation• carries amino acids to mRNA• carries anticodon to mRNA• translates a codon of mRNA into an amino acid
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rRNA Molecules
Ribosomal RNA (rRNA) –• provides structure and enzyme activity for ribosomes
• ribosomes are necessary for protein synthesis
• Where in the cell are ribosomes manufactured?
Figure from: Alberts et al., Essential Cell Biology, Garland Press, 1998
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Mutations
Mutations – change in genetic information
Result when • extra bases are added or deleted• bases are changed
May or may not change the protein
Repair enzymes usually correct mutations
This single point-mutation causes sickle cell disease!
Figure from: Hole’s Human A&P, 12th edition, 2010
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Mutations
Recall that the 3-D structure of proteins are dependent, ultimately, upon the primary (linear) sequence of the protein.
So, a change in a single amino acid of a protein may affect the subsequent levels of protein structure.
Would such a mutation have any advantage?
What if only one allele of the -globin gene was affected?
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Central Dogma of Molecular Biology
Applicable to all cells from bacteria to humans.
Genetic information flows from:
DNA RNA Protein
(Central Dogma)
Transfer of information into protein is irreversible
Figure from: Alberts et al., Essential Cell Biology, Garland Publishing, 1998
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Overview of Protein SynthesisFigure from: Hole’s Human A&P, 12th edition, 2010
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Transcription
Recall that the nitrogenous bases in nucleic acids can hydrogen bond to each other in a complementary fashion.
A T (U) and G C
Thus, one strand of a nucleic acid (a gene) can serve as a template for the generation of a new strand.
Note that transcription takes place in the NUCLEUS of the cell.
The generation of mRNA (nucleic acid) from DNA (nucleic acid)
*
Figure from: Hole’s Human A&P, 12th edition, 2010
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Transcription
Figure from: Martini, Human Anatomy & Physiology, Prentice Hall, 2001
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TranscriptionGenes are directional; the template for making a protein is located on ONE strand (usually) of DNA
DNA of chromosome
Figure from: Alberts et al., Essential Cell Biology, Garland Publishing, 1998
REMEMBER:
Template Transcribed (“Template strand is transcribed”)
Coding Codon (“Coding strand LOOKS LIKE the codon”)
Coding
Template
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Eucaryotic Genes Are Not Continuous
Figure from: Alberts et al., Essential Cell Biology, Garland Publishing, 1998
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mRNA Modification
Figure from: Alberts et al., Essential Cell Biology, Garland Publishing, 1998
Newly made eukaryotic mRNA molecules (primary transcripts) undergo modification in the nucleus prior to being exported to the cytoplasm.
1. Introns removed2. 5' guanine cap added3. Poly-A tail added
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Translation
• How does the cell convert (translate) the symbols of nucleic acid into the symbols of amino acids?
• Does this happen directly, or is there some intermediate, e.g., a key of some sort?
Generation of a polypeptide (amino acids) from mRNA (nucleic acids) in the cell’s cytoplasm
*
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The Genetic Code
1. There are a TOTAL of 64 possible codons…2. Of these 64 codons, 61 are actually used to code for amino acids3. Notice that more than one codon may correspond to a specific amino acid.
Table from: Hole’s Human A&P, 12th edition, 2010
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The Genetic Code
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Overview of Translation
Transfer RNAs (tRNA) function as ‘adapters’ to allow instructions in the form of nucleic acid to be converted to amino acids.
Figures from: Martini, Anatomy & Physiology, Prentice Hall, 2001
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Attachment of Amino Acids to tRNA
How is the correct amino acid associated with its corresponding tRNA?
Enzymes! (aminoacyl-tRNA synthetases)
There are 20 synthetase enzymes; one for each amino acid
‘Charged’
Figure from: Alberts et al., Essential Cell Biology, Garland Publishing, 1998
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Translation
Figure from: Martini, Human Anatomy & Physiology, Prentice Hall, 2001
Ribosomes in the cytoplasm are critical to the generation of proteins during translation
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Translation
Figure from: Martini, Human Anatomy & Physiology, Prentice Hall, 2001
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Translation
Figure from: Martini, Human Anatomy & Physiology, Prentice Hall, 2001
One of three possible STOP codons (UGA, UAG, UAA)
So, what is the set of ‘rules’, the key, by which a particular codon corresponds to a particular amino acid (aa) called?
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Review of Protein SynthesisFigure from: Hole’s Human A&P, 12th edition, 2010
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The Fate of Proteins in the Cell
• Breakdown of proteins regulates the amount of a given protein that exists at any time.
• Each protein has unique lifetime, but the lifetimes of different proteins varies tremendously.
• Proteins with short life-spans, that are misfolded, or that become oxidized must be destroyed and recycled by the cell.
Enzymes that degrade proteins are called proteases. They are hydrolytic enzymes.
Most large cytosolic proteins in eukaryotes are degraded by enzyme complexes called proteasomes.
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Find the AMINO ACID SEQUENCE that corresponds to the following gene region on the DNA:
Template -> G A T T G A A T C
Coding -> C T A A C T T A G
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The Genetic Code (Codon Table)
Do the Extra Credit…only if you want to do well on exam!
Table from: Hole’s Human A&P, 12th edition, 2010
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Review
• A gene is a stretch of DNA that contains the information to make protein
• A genome contains all the genes of an organism
• The Genetic Code is the method used to translate a sequence of nucleotides of DNA into a sequence of amino acids
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Review• The genetic information of the cell is
contained in its nuclear DNA– DNA is packaged in the nucleus into chromatin
(DNA plus histone proteins)– DNA is a anti-parallel, double-stranded helical
polynucleotide containing deoxyribose– Four bases are used in DNA:
• Purines (double ring): Adenine (A), Guanine (G)• Pyrimidines (single ring): Cytosine (C), Thymine (T)
– A pairs with T using two (2) hydrogen bonds– G pairs with C using three (3) hydrogen bonds
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Review
• RNA is a polynucleotide with important differences from DNA– Uses Uridine (U) rather than Thymine (T)– Uses the pentose sugar, ribose– Usually single-stranded
• There are three important types of RNA– mRNA (carries code for proteins)– tRNA (the adapter for translation)– rRNA (forms ribosomes, for protein synthesis)
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Review
• DNA replication– During interphase– Creates an identical copy of the genetic information– Semi-conservative replication (one old, one new
strand)– Uses DNA polymerase
• Matches complementary bases with template
• Replication forks
• Error-correcting capability
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Review• Mutations are errors in the genetic material
(DNA)– May affect the end-product, i.e., the protein– Vary in type and severity– Must become ‘fixed’ in the cell to be passed to
future generations (sickle cell disease)
• Mutations at the chromosomal level may be caused by– Deletions– Translocations– Extra copies of chromosomes
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Review
• Genetic information flow in the cell is from DNA RNA Protein
(Central dogma of molecular biology)• Transcription generates mRNA from DNA• Translation generates polypeptides (proteins) from
mRNA using tRNA and ribosomes• The genetic code is the set of specific instructions
for translating nucleic acid information into proteins
• The life-span of proteins in the cell is limited by degradation by proteases in complexes called proteasomes.