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MMBIO Quiz 9
| Question | Answer |
|---|---|
| prokaryotic transcription | 1 main polymerase RNA polymerase binds to DNA directly no nucleu |
| eukaryotic transcription | 3 RNA polymerases with more subunits nucleus exports mRNA to cytoplasm mRNA processin more genes=more complicated |
| RNA pol I | pre-rRNA and large rRNA |
| RNA pol II | pre mRNAs, snRNAs |
| RNA pol III | tRNA, rRNA |
| we can use GFP (reporter gene) to measure expression by cloning the promoter in a vector | very green - high promoter expression light green - low promoter activity no green - no promoter activity |
| reporter assay | use luciferase to measure enhancer/promoter activity |
| what TF recruit RNA pol to bind? | ChIP assay |
| what negative regulators not allow RNA pol to bind? | ChIP assay |
| what regulatory regions exist in the DNA sequence? | site-directed mutagenesis |
| what drugs can activate/block regions? | study GFP production levels |
| transfection | introduction of foreign DNA into eukaryotic cells |
| basal RNAPII complex | group of proteins that bind to promoter regions and brings RNAPII to begin transcription |
| core promoter (-40 to +40) | TATA box -31 to -26 InR -2 to +4 DPE +28 to +32 |
| minimal promoter | only has 2 of the 3 core promoter elements, rare that all 3 are together you must always have InR though |
| TBP | TATA binding protein binds to DNA to initiate transcription is a part of TFIID |
| initiator | YYANA/TYY first base transcribed is often A |
| INITIATION order of basal complex | TFIID (with TBP) binds to DNA on TATA box TFIIA stabilized binding TFIIB binds complex and recruits RNAPII TFIIF brings in RNAPII TFIIE recruits TFIIH TFIIH has helicase and kinase activity to open the template and phosohrolaytes RNAPII |
| TFIIH phosphorylates the ___ on the RNAPII which triggers transition to elongation | CTD (C-terminal domain) |
| kinase | enzyme that adds phosphate groups onto molecules |
| RNAPII delays at pause sites | arrest/proofreading |
| 4 DNA-binding motifs that fit inside the grooves | Helix turn Helix Helix loop Helix Zinc Fingers Leucine Zippers |
| transcription full steps | chromatine is remodeled/loosened activator proteins bind to regulatory promoters DABFEH and RNAPII bind to TATA box via TBP DNA is unwound RNAPII moves along DNA< synthesizes RNA Mediator - additional rounds mRNA processing termination and export |
| regulatory promoters | where transcription activator proteins bind to assist TFIID to recruit the basal complex |
| enhancers | can be anywhere in any orientation |
| activator proteins | activation domain (interact with RNAP and Mediator complex) DNA binding domain |
| coactivators | do not bind to DNA directly help TFs interact with basal TFs at promoter |
| DNA looping | allows distal regions to have an effect on promoters allows enhancers to work everywhere |
| Mediator complex | interacts with activators on regulatory promoters gives stronger signal to recruit RNAPII increases rate/efficiency of RNAPII assembly |
| 4 methods to identify which DNA sequences and proteins are involved in initiation (2 of 4) | DNA affinity chromatography - specific ones attach to beads ChIP assay |
| 4 methods to identify which DNA sequences and proteins are involved in initiation (2 of 4) | promoter deletions - if expression is way lower after deletion, it is a promoter linker scanning mutagenesis - like deletion but replacing with non functional regulators to preserve distance |
Created by:
anyasalmon