Regulation Of Gene Expression Flashcards
Gene expression results in the production of
Either RNA or protein
Describe each of the following kind of genes:
- Housekeeping genes
- Regulated genes
- Involved in basic cellular function, constitutively expressed and not regulated
- Required only in certain cell types/ certain conditions; subject to various control mechanisms that will determine if these genes will be expressed
Regulation of gene expression in pro:
- 2 regulatory molecules and their function
- 2 types of operons and their function
- Repressors and activators: suppress/increase transcription of a gene
- Inducible - transcription is usually OFF but can be stimulated
Repressible- transcription is usually ON but can be inhibited
Lac operon:
- Prokaryotes or eukaryotes?
- Inducible or repressible?
- When would it be induced
- What type of bacteria uses this
- Prokaryotes
- Inducible (only inducible one we need to know)
- When glucose is absent but another sugar is present
- E. coli
Lac operon
- When only glucose is present, lac operon is on or off?
- Repressor protein is encoded by? Where is it? What enzyme does it block?
- Glucose inhibits what enzyme? What would this cause?
- Off
- LacI gene; always present and bound to the operator; blocks RNA polymerase
- Adenylyl cyclase -> no cAMP, cannot form CAP/cAMP complex, cannot initiate transcription
Lac operon continued:
- If only lactose is present, is it on or off?
- When glucose is absent what happens
- When lactose is present, what is produced? What is it and what does it do
- On
- Adenylyl cyclase makes cAMP, CAP/cAMP complex forms, binds to CAP binding site, RNA polymerase can initiate transcription
- Allolactose (small amount), isomer of lactose produced from lactose, binds to the repressor and prevents it from binding to the operator (so gene is not blocked)
Lac operon continued:
- When both lactose and glucose are present, lac operon is on or off?
- Is repressor active or inactive?
- Then why can’t transcription be initiated?
- Off
- Inactive
- CAP site is empty because glucose is present
Eukaryotes are different because they have gene expression regulation at 5 different levels. Give an example
- Transcriptional
- Posttranscriptional
- Translational
- Posttranslational
- Epigenetic
- PEPCK
- Alternative splicing/mRNA editing
- EIF-2
- Modifications of polypeptide chain
- Modifications of DNA
Transcriptional control:
- Regulatory sequences of DNA are usually embedded __
- Define cis-acting elements
- Why are regulatory sequences of DNA called cis-acting
- Interact with? Which are?
- Trans-acting regulators are __
- Noncoding region of the genome
- DNA sequences bound by trans-acting regulatory molecules (enhancers)
- They influence expression of genes only on the same chromosome
- Regulatory molecules (transcription factors) which are trans-acting regulators
- Proteins
Binding of trans-acting regulators to DNA is achieved by one of which 3 things?
Zinc finger, leucine zipper, helix-turn-helix in protein
- Define enhancer
- Can be located?
- When can they act in a tissue specific manner?
- DNA sequences that increase rate of initiation of transcription
- Close or far from the gene, or even within intron regions of other chromosomes (typically on same chromosome tho)
- If the DNA binding proteins (transcription factors) are only present in certain tissues
- How can enhancers be brought close to the basal promoter
- Transcription factors are cis or trans?
- Transcription factors interact with RNA polymerase II to stabilize formation of __ and recruit?
- By bending the DNA molecule
- Trans
- Initiation complex and recruit chromatin modifying proteins (histone acetylase)
PEPCK gene expression is induced by __ which diffuses into ?
Cortisol; hepatocyte
- Function of alternative splicing
2. Give an example
- To make tissue specific isoforms of proteins from the same pre-mRNA
- Tropomyosin
- MRNA editing is what type of control?
2. Example: how does apo B100 get changed to apo B48
- Post-transcriptional
2. CAA codon for glutamine is deaminated to U (UAA) which is a stop codon - makes protein shorter (only gets 48% of)
MRNA stability:
- RNAi (interference) reduces gene expression by either of what 2 things
- Plays a role in what 3 processes
- Has huge __ potential
- Repressing translation and increasing degradation of specific mRNAs
- Cell proliferation, differentiation, and apoptosis
- Therapeutic
- RNAi is mediated by? How many base pairs does it have?
- MiRNA acts as ?
- Works with RISC-RNA-induced silencing complex to do what?
- What can also trigger RNAi?
- MicroRNA (miRNA) ~20-22 bp
- Guide strand (contains complementary sequence)
- Block translation
- SiRNA
- MicroRNA (miRNA) ~20-22 bp
Translational control:
- Phosphorylation of eIF-2 inhibits its function how?
- Name some things that would activate kinases to phosphorylate eIF-2 (4)
- By inhibiting GDP-GTP exhange thus inhibiting translation at the initiation step
- Amino acid starvation, heme deficiency, presence of double stranded DNA, accumulation of misfolded proteins in the rER
- Why is cleavage/trimming necessary for many proteins? Example
- Protein degradation is done by
- To become functionally active; protein digestion enzymes
2. Ubiquitination
Euchromatin vs heterochromatin
Euchromatin= loose, accessible for transcription
Heterochromatin= tightly packed, inaccessible
Epigenetic regulation:
- What are CpG islands
- Methylation of DNA/histones causes? Genes are expressed or no?
- Histone acetylation causes? Genes are expressed or no?
- Regions in DNA rich in CG that are prone to modifications
- Nucleosomes tightly pack together; genes are not expressed
- Nucleosomes are loosely packed; genes are expressed
- What are transposons
- Direct vs replicative movement
- Transposition contributes to __
- Transposition is associated with what 3 diseases/conditions?
- Mobile segments of DNA that move in a random manner from one site to another one same or different chromosome
- Direct=cuts out and inserts transposon at a new site; replicative= transposon is copied and inserted elsewhere while original remains in place
- Structural variation in the genome
- Hemophilia A (rare), duchenne muscular dystrophy, antibiotic resistance in bacteria