Unit 7: Gene Expression
Describe the mechanisms by which genetic information flows from DNA to RNA to protein.
Transcription: The process by which the information stored in DNA is copied into RNA
Occurs in nucleus for eukaryotes
The DNA double helix unwinds at the gene location
RNA polymerase enzyme reads the DNa template strand 3’ to 5’ direction
Complementary RNA nucleotides are added to form mRNA
The mRNA molecule is processed (in eukaryotes) through steps like RNA splicing
The mature mRNA exits the nucleus and enters the cytoplasm
Translation: The process by which the mRNA is used as a template to synthesize proteins
Occurs in the cytoplasm and involves ribosomes
The ribosome binds to the mRNA at the start codon (usually AUG)
tRNAs bring specific amino acids to the ribosome
The ribosome reads the mRNA codons, and matching tRNAs deliver their amino acids
Amino acids are linked together to form a growing polypeptide chain
The process continues until a stop codon is reached
Transcribe and translate a segment of DNA to determine the amino acid sequence of the protein created.
Given DNA template strand
5' - TACGCAATGGCCTAA - 3'
Transcription into mRNA:
mRNA is complementary to the template DNA strand, and uses U instead of T
5' - AUGCGUUACCGGAUU - 3'
Translation:
Using the genetic code to convert each codon group (3 nucleotides per codon) into its corresponding amino acid
mRNA codons and their corresponding amino acids
AUG - Methionine (Start codon)
CGU - Arginine
UAC - Tyrosine
CGG - Arginine
AUU - Isoleucine
Final protein sequence
Methionine - Arginine - Tyrosine - Arginine - Isoleucine
Met-Arg-Tyr-Arg-Ile
Describe how the phenotype of an organism is determined by its genotype.
An organism’s genotype is the set of genes it carries, which contains the instructions for its growth and development
The phenotype is the observable expression of these genetic instructions
Some factors
Direct influence
Gene interactions
Environmental factors
Dominance and recessiveness
Epigenetic factors
Explain the types of post-transcriptional modifications to eukaryotic mRNA and its purpose within the cell.
5’ Methyl Cap
Modified guanine nucleotide added to 5’ end
Helps ribosome start translation
Poly-A-Tail
Many (100-200) adenine nucleotides added to the 3’ end
Protection from degradation in cytoplasm (lifespan of mRNA)
Splicing
Introns removed, exons joined
Spliceosome
Exons code for protein
Describe the various types of mutations.
Mutations: Changes in the genetic material of an organism
Point Mutations: Changes to ONE nucleotide in the DNA sequence
Substitutions: one base is replaced with another
Transitions: A purine is replaced by another purine, or a pyrimidine by another pyrimidine
Transversions: A purine is replaced by a pyrimidine, or vice versa
Insertions: An extra base is added to the DNA sequence
Deletions: A base is removed from the DNA sequence
Effects of Point Mutations
Silent mutations: No change in the amino acid sequence
Missense mutations: Result in a different amino acid
Nonsense mutations: Create a premature stop codon
Frameshift Mutations: When the number of nucleotides inserted or deleted is not a MULTIPLE OF THREE
Leads to a completely different amino acid sequence or premature termination of the protein
Causes of Mutations
Spontaneous errors during DNA replication
Exposure to mutagens, such as UV radiation, X-rays, or certain chemicals
Explain how changes in genotype may result in changes in phenotype (Hint: Answer should include description of protein folding).
Mutations and Protein Folding: They can alter the amino acid sequence of proteins, which in turn can affect protein folding
Stability changes
Mutations can increase or decrease protein stability
Folding defects
Accumulation of folding defects can reduce a protein’s ability to evolve a new phenotype
Functional changes
Alterations in protein folding can lead to changes in protein function.
Protein Abundance and Evolvability: The abundance of a protein can influence its evolvability
Low Expression
Proteins expressed at lower level may be more evolvable toward new phenotypes
High Expression
Highly expressed proteins may be less evolvable, as they can tolerate more genetic variants that cause folding defects
Explain how alterations in DNA sequences contribute to variation that can be subject to natural selection.
Already did it
Explain how the binding of transcription factors to promoter regions affects gene expression and/or the phenotype of the organism.
Activation and repression: TFs can act as activators or repressors of gene transcription
Activators enhance the recruitment of RNA polymerase and other general transcription factors to the promoter
Repressors can block the binding of these essential components, reducing or preventing transcription
Describe the regulation of gene expression via operons in prokaryotes.
Structure of an Operon
Promoter
A DNA sequence where RNA polymerase binds to initiate transcription
Operator
A regulatory sequence near the promoter where regulatory proteins binds to the promoter
Structural genes
The genes encoding proteins with related functions
Regulatory Mechanisms
Induction
Inducers are small molecules that can bind to repressors to cause a conformational change that prevents the repressor from binding to the operator
This allows RNA polymerase to transcribe the genes
Repression
Repressor proteins bind to the operator, WHICH THEN blocks RNA polymerase from transcribing the genes
In the absence of an inducer, the repressor is active and gene expression is inhibited
Activation
Activator proteins can enhance transcription by binding to specific DNA sequences
They facilitate the binding of RNA polymerase to the promoter
EXAMPLE: THE LAC OPERON
The Lac Operon in E. coli is an example of operon regulation
It contains genes for lactose metabolism (lacZ, lacY, lacA)
In the absence of lactose, a repressor protein binds to the operator, therefore preventing transcription
IF AND WHEN LACTOSE IS PRESENT it acts as an inducer by binding to the repressor and allowing transcription to occur
Contrast inducible and repressible operons.
Inducible
Typically off by default
Activated by the presence of an inducer molecule
Often associated with catabolic pathways
Example: LAC OPERON activated by LACTOSE
Repressible
Typically on by default
Deactivated by the presence of a co-repressor molecule
Often associated with anabolic pathways
Example: TRP operon repressed by TRYPTOPHAN
Describe various mechanisms that allow for cell differentiation of cell specialization in multicellular eukaryotic organisms.
Differential gene expression: Cells selectively activate or repress specific gene leading to the production of proteins that determine cellular structure and function
Transcription factors: Proteins that bind to specific DNA sequence to control gene expression
Chromatin modifications: Changes in DNA packaging that affect gene accessibility
Epigenetic mechanisms: Such as DNA methylation and histone modifications
Asymmetric cell division: During cell division, regulatory molecules are unevenly distributed between daughter cells, resulting in distinct developmental fates
Cytoplasmic determinants: Molecules inherited unequally by daughter cells
Cell signaling: External signal that influence cell fate decisions
Explain the connection between the regulation of gene expression and phenotypic differences in cells and organisms.
Explain the life cycle of viruses and the role of reverse transcriptase in allowing viruses to evade the immune system and treatment.
Life cycle of virus
Attachment: The virus binds to specific receptors on the host cell surface
Penetration: The virus enters the host cell
Uncoating: The viral genome is released inside the host cell
Biosynthesis: The virus hijacks the host cell machinery to replicate its genetic material and produce viral proteins
Assembly: New viral particles are formed
Release: The newly formed viruses exit the host cell
Reverse transcriptase
Genome conversion: Reverse transcriptase converts the viral RNA genome into DNA, enabling integration into the host cell’s genome
High mutation rate: The enzyme lacks proofreading activity, leading to frequent mutations that help the virus evade immune recognition and develop drug resistance
Rapid evolution: The high mutation rate allows the virus to quickly adapt to antiviral drugs, making treatment challenging
Latency establishment: By integrating viral DNA into the host genome, reverse transcriptase enables the virus to establish latent infections, evading immune detection
Genetic variability: The error-prone nature of reverse transcriptase contributes to HIV-1’s genetic diversity, further complicating immune recognition