Block 5 Flashcards
What happens during the oxidation of bases?
Generation of strand interruptions from reactive oxygen species
What happens during G1 to proceed thru the cell cycle?
Growth factors induce expression of cyclin D. Cyclin D associates with CDK4/6 and they inactivate Rb
What are the steps to extrinsic apoptosis?
- fas ligand on killer lymphocyte binds fas receptor (juxtacrine signaling) 2. oligomerization of receptor and recruitment of death domain with adaptor proteins. 3. adaptor proteins contain death effector domains that recruit and activate initiator procaspase 8 4. caspase 8 is cleaved and activates effector caspases
What is the function of E2F in G1?
E2F induces expression of DNA replication enzymes, cyclin D, cyclin E, and cyclin A
What causes HNPCC?
Mutations in MMR proteins MSH2/ MLH1
What happens when JAK proteins are activated?
Create binding sites for STAT proteins
How are origins of replication “licensed” during G1?
Sequential assembly of origin replication complex, CDC6, CDT1, and mcm helicases
True or false: eukaryotic DNA polymerases have 5’ to 3’ exonuclease activity?
FALSE. Require 5’ flap endonucleases
What mutagens cause missing bases?
acid and heat depurination
What are the steps to autophagy?
autophagosome engulfs portions of the cytoplasm. Autophagosome becomes acidified and forms autolysosome
How does AZT work?
T base analog for fighting HIV
What is the function of DNA PKCs + ku 70/80 system?
Fix double strand breaks
What is the unfolded protein response?
Triggering of apoptosis due to unfolded proteins in ER via caspase 12.
Name and explain the two prokaryotic DNA replication models
- Theta model: DNA unwinds @ origin in replication bubble. Leading strand in direction of fork with lagging strand opposite 2. Rolling circle: continuous synthesis @ 3’ end and 5’ end displaced
Why are telomeres needed?
At the end of chromosomes, there are no places for RNA primers to start the last Okazaki fragment
What are the symptoms and causes of trichothiodystrophy?
Defects in NER cause light sensitivity, premature aging, ichtyosis, and sulfur-deficient brittle hair
What is the function of BRCA2?
Regulates homologous recombination
What types of tumor-suppressor proteins can be mutated in cancer?
receptors/ signal transducers in inhibiting cell cycle (TGF-B), cell cycle inhibitors, check-point control proteins (p53), pro-apoptotic proteins, DNA repair enzymes
What are the functions of cyclins?
Regulate activity of CDKs and regulate substrates that CDKs can phosphorylate. Cyclin+ CDK = master regulators
What happens during prophase?
Chromosome condensation (condensins) and spindle assembly begins
What mutagens cause deletions/ insertions?
intercalating agents (Acridines)
How does MMR know which strand is the parent strand?
Parent strands are methylated while new daughter strands have not had time to be mehtylated
What is the function of cyclinE-CDK2 at the end of G1?
Fully activates E2F and assembles preinitiation complexes on chromosomes
What is the effect of thermal disruption?
Due to elevated temperatures. Increases rate of depurination and single strand breaks.
What is the function of mismatch repair (MMR)?
Correction of mismatches of normal bases via NER and BER proteins
What is the function of non-homologous end joining?
“quick and dirty” end repair that causes DELETION of DNA or other mutations because it joins dissimilar DNA sequences.
What forms the apoptosome and what is its function?
Cytochrome C + Apaf 1. Cleave and activate procaspase 9
What is the function of phosphatidyl serine in apoptosis?
Normally is located on the inner leaflet. During apoptosis, i is flipped out to outer leaflet to provide signal for phagocytosis
What mutagens cause altered bases?
ionizing radiation and alkylating agents
What happens if there is sustained cyclin B expression in a cell?
Cell becomes quiescent
What percentage of breast cancer is inherited? What percent of those inherited cases are linked to BRCA1 or BRCA2?
5-10% inherited; 60-80% inherited cases linked to BRCA1/2
What are some features of Okazaki fragments?
1000-2000 NT in prokaryotes and 100-200 in eukaryotes. Synthesis of each fragment on the lagging strand begins @ the replication fork.
What happens when TGF-B binds the TGF-B receptor?
3 receptors oligomerize and phosphorylate Smad proteins to activate them, causing a conformational change.
How is p21 turned on and what pathways is it involved in?
Turned on via p53. Involved in DNA damage checkpoints
Describe Ras in its inactive state
Bound to GDP and usually bound to the membran
What medication is being used to treat CML?
Gleevac
What is Rbs function when it’s active?
Rb is a tumor suppressor and binds E2F
What happens during telophase?
NE reforms and cytokinesis occurs.
What type of mutations is seen in oncogenes?
Gain of function
What is the effect of Ras mutations in cancer?
Reduction of GTPase activity so more of the protein is found in the GTP bound active form
What effect can epinephrine have on heart muscle?
epinephrine activates GPCR and GPCRs activate MAPK which can lead to cardiac hypertrophy.
What happens when a ligand binds the cytokine receptor?
JAK lip kinase is activated via phosphorylation
What is the relationship between cancer cells and stromal cells?
Cancer cells influences stroma and stromal cells influence tumor growth. Ex melanoma cells release PDGF to stimulate fibroblasts in stroma. Fibroblasts secrete IGF-2 survival factor
True or False: homologous DNA recombination occurs between dissimilar sequences?
FALSE: only occurs between highly similar sequences
What is the significance of the RTK pathway in small cell lung cancer?
Extracellular EGF receptors are cleaved and dimerize without ligand binding to be constituently active.
What are some properties of mesenchymal cells after EMT?
gain of fibroblast shape, motility, invasiveness, mesenchymal gene expression, protease secretion, vimentin expression, PDGF recetor expression, fibronectin expression, etc.
What cyclin accumulates at the end of G1?
cyclin E that will bind to CDK2
What is the function of recombination enzymes?
Recognize and bind inverted repeats in recombination sites to form synaptic complexes.
What are the steps to metastasis?
- primary tumor formation 2. localized invasion 3. intravasation into blood stream/ lymph 4. transport thru circulation 5. arrest in microvessels of organs 6. extravasation 7. micrometastasis formation (needs good stroma) 8. colonization and formation of macrometastasis
What is the function of cyclin B in the M phase
Phosphorylates lamins, condensins, microtubule associated proteins, APC/C
What are the properties of prokaryotic DNA replication?
Semiconservative, bidirectional, one origin per chromosome, semidiscontinuous, RNA primed
What are the properties of eukaryotic DNA replication?
Semiconservative, bidirectional, multiple origins/ chromosome, semidiscontinuous, RNA primed, replication of telomeres, nucleosome assembly
How are mobile genetic elements related to brain function?
Can contribute to brain diversity, but are linked to schizophrenia
What enzymes fill in the gaps of BER?
DNA pol B fills in gap and ligase joins strand
How do the autophagy and apoptosis pathways cross-talk?
Formation of autolysosomes can block caspase activation or caspase activation can block autophagosomes
What is the function of Raf in the MAPK pathway?
Raf phosphorylates and activates MEK
What is the function of BRCA1 in relation to p53, Wee 1, and CDC 25?
BRCA1 is a scaffold protein that enhances p53 phosphorylation and stabilization during DNA-damage induced cell cycle checkpoints. Also regulates Wee1/ CDC25 in G2/M checkpoints.
What happens to Smad proteins after they are activated and undergo conformational change?
Smad proteins dimerize and enter the nucleus to bind DNA and NEGATIVELY regulate growth
What does B-catenin do if not degraded?
Associates with TCF and activates genes for gene expression in the nucleus
What is the effect of aflatoxin?
Procarcinogen that can form bulky adducts and cause DNA mutations
How is Rb inactivated?
Hyperphosphorylation by cyclin D-CDK4/6
What is the function of Ras in the RTK pathway?
Ras activates the Mitogen Activated Protein Kinase Pathway by activating Raf
How are CDKs activated?
Cyclin binds and causes a conformational change and ATP binding. Phosphorylation on T-loop is exposed so it can be phosphorylated by CAK.
What is the function of cdc6/ cdt1?
Recruited after ORC binds and recruit mcm2-7 helicases. They are phosphorylated by CDKs
What are the effects of radiation therapy?
double strand breaks and reactive oxidative species generation to induce apoptosis.
What are the effects of alkylating agents?
Prevent DNA synthesis and transcription via blocking DNA separation, induce mutations, cause inter & intrastrand ligations and DNA cross-links
How does overexpression of the cyclin D gene contribute to cancer?
cell becomes extremely sensitive to growth factors and pushes faster thru G1. (43% head and neck and 50% breast cancers)
Explain the disease mechanism in Chronic Myelogenous Leukemia (CML)
Chromosomal translocation where the abl kinase is fused to the bcr gene. Change in substrate specificity of abl kinase
What kind of binding domain does Grb2 have and what is its function in the RTK pathway?
Grb2 has SH3 domains to bind to SH2 and bind SOS
What happens to B-catenin in the absence of Wnt ligand?
Targeted to the destruction complex, phosphorylated, ubiquitinated, and degraded
What do dimerized STAT proteins do in the nucleus?
Bind DNA directly and activate transcription
What are the three methods of mutagenesis?
- Endogenous/ spontaneous 2. Genetic defects 3. Exogenous mutations
What are the effects of intercalating agents?
Slide in between bases of the DNA duplex and disrupt replication and transcription. Usually have planar ring structure.
What is the significance of the Wnt pathway in colon cancer?
APC is usually mutated so it cannot form the destruction complex.
What comprises a “bead” of packed eukaryotic DNA?
8 histone proteins with 146 bp DNA
What is the function of the tau subunit in DNA polymerase III?
interacts with helicase and holds DNA polymerase subunits together
What is the function of ribonuclease H1?
Nuclease specific for RNA in RNA/ DNA hybrids to excise primers. Does NOT remove last ribonucleotide from initiator DNA
What are some outcomes of site-specific recombination?
gene/ DNA integration, excision, inversion
What does base excision repair fix?
Fixes only base (purine or pyrimidine)
What are the symptoms and causes of Xeroderma pigmentosum?
Hereditary defect in proteins for NER important for repair of UV-induced DNA damage. Symptoms: severe light sensitivity and increased skin cancer
Where was the Rb protein first found?
identified in patients with hereditary retinoblastoma and those cells were found to have lost both alleles of Rb
Which cyclins bind CDK2?
Both cyclin E and cyclin A. Direct CDK2 to different substrates
What is the effect of UV-A?
Causes free radical generation and INDIRECT DNA damage
How does Ras dissociate from Faf?
GTP hydrolysis
What happens when microtubule associated proteins are phosphorylated?
Form the mitotic spindle
What happens without the survival signal in anoikis?
AKT is not activated so BAD is not phosphorylated. BAD induces cytochrome C release and caspase 9 activation
What is the function of 6-mercaptopurine and 6-thioguanine?
Interfere with purine synthesis
What is the role of DnaC in prokaryotic DNA replication?
Helps to load DnaB helicase onto DNA
What happens during prometaphase?
Disassembly of the NE, complete spindle assembly, and chromosomes begin to assemble @ both poles
What is the common pathway for TGF-B and Cytokine signaling?
Activation of the receptor causes phosphorylation of proteins that can translocate to the nucleus and regulate gene txn. Less intermediate steps than RTK
True or false: mobile genetic elements require specific sequence homology?
FALSE: can insert into “any or many” DNA sequences
How do inhibitory smads block TGF-B expression?
They are induced by TGF-B (negative feedback loop) and block Smad phosphorylation.
What happens to STAT proteins when the bind to the JAK kinase?
STAT proteins dimerize via SH2 domains and translocate into the nucleus
In what step of cancer therapy are we most lacking?
Developing appropriate drugs and delivery in clinical studies
What are some examples of intercalating agents?
Actinomycin D, aflatoxins, echinomycin
What is the function of AP endouclease in BER?
removes sugar phosphate
What types of mutations activate NER?
Bulky lesions (ex. thymine dimers)
What happens during anaphase?
Physical separation of chromosomes, sister chromatids move to opposite poles, poles move apart.
What happens when C:G is deaminated?
Changed to T:A or C to T point mutation. Can’t be repaired
How much of the genome is non-coding?
98%- 50% repeating sequences & 50% unique sequences
What is the function of Fen-1?
5’ flap endonuclease that removes last ribonucleotide from initiator DNA
How does the SHP1 phosphatase terminate the cytokine signaling pathway?
Inactivates JAK by dephosphorylation