Control of Gene Expression (Ford L1) Flashcards
What does a DNA control element do?
Acts locally (cis)
Binding of trxn factors to a DNA control element regulates the gene the control element is associated with.
What are two broad classes of gene expression regulation?
- Control Elements - act locally - cis
2. Trxn’l Activators and Repressors - act at a distance - trans
Give three examples of DNA control elements.
- TATA box
- Promoter Proximal Elements
- Enhancers
Where is the TATA box (initiator seq.) and what does it do?
23-35 bp upstream of trxn start site 20 bp long determines site of trxn initiation directs/binds RNA pol II binds general trxn factors (TF II ?)
What are basal trxn factors?
Base trxn factors that bind to all promoters to recruit in the rest of the trxn machinery & RNA pol II
What are Promoter Proximal Elements?
located 200 bp upstream of trxn start site
8-20 bp long
Help to regulate trxn
are bound by cell specific factors (or by factors found in all cell types)
What would happen if you moved a promoter proximal element kilobases upstream (or downstream)?
Would not function properly.
PPE’s need to be relatively close to the gene they are regulating (200 bp)
What are enhancer elements?
can be 200 - 10kb up or down stream of start site OR within an intron
is 100-200 bp long but made up of many small (8-20bp) control elements (binding sites for different factors).
are often cell-type specific
Give three examples of diseases arising from mutations in DNA control elements.
- Beta-Thalassemia or (gamma-delta-beta) Thalassemia
- Hemophilia B Leyden
- Fragile X-Syndrome
Give two examples of Transcriptional Activators / Repressors
- Sequence specific DNA binding proteins
2. Co-factors - bind to the DNA binding proteins (not to the DNA itself)
How do sequence specific DNA binding proteins do their job?
They bind to a specific sequence (DNA control element !) 6-8bp longInsert alpha-helix into MAJOR groove of DNAContact between aa side chains and DNA bases
Distinguish between major and minor groove binding.Give an example of a trxn protein that binds the minor groove
Major groove binding is very sequence specific. Minor groove binding will distort the DNA more.The TBP (TATA binding protein) binds to the minor groove using the beta sheet.
TATA box and bends it to a right angle, inserts aa’s in between base pairs, thus partially unwinding DNA. So it has a small amount of melting activity.
What is modularity in sequence-specific DNA binding proteins?
have specific domains that do specific things. if you swap out a module from one gene into another gene, the second gene will gain the activity of the inserted module
Name 3 specific homeodomain proteins (genes)
- Hox family - regulates anatomical body plan (antennapedia mutation)
- Pit1 - pituitary specific - growth hormone regulation
- Msx - neural crest cells responsible for proper craniofacial development
What is the general geometry of a homeodomain sequence specific DNA binding protein? Where does it bind?
Helix - Turn - Helix
Binds to the major groovetwo alpha helices connected by a turn of DNA, running antiparallel to one another
A third alpha helix connected by a turn of DNA, running perpendicular to the first two