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Lecture 9

BIO SCI 99 Lecture 9: Bio 99 Lec 9


Department
Biological Sciences
Course Code
BIO SCI 99
Professor
Tatianne
Lecture
9

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LECTURE 9
EUKARYOTIC TRANSCRIPTION
TATA box
Int: initiator sequence
Phosphorylation of CT domain during initiation
Dephosphorylation after elongation
Pol I: rRNA
UBF: upstream binding factor
SL1: protein complex that includes the TATA-binding protein
Pol III: tRNA, 5S rRNA
Promoter sequences within the coding sequences
Major groove = more
Facilitate interaction and stop at certain sequences
Txn factors tend to function as dimers
Modular Txn Factor Domains
DNA binding domain
Activation/Repressor domain
Bind RNA polymerase
Other txn factors
Yeast 2 Hyrbid:
each txn factor will ha e bind site and DNA activation site
Bait protein - Gal4 DNA binding domain fused to protein X
Prey protein - Fal4 activation domain fused to protein Y (DNA library)
Txn Factor Domains - Yeast 2 Hybrid
A gal4 yeast strain ___ grow on galactose containing media - WILL NOT
Cannot use galactose as source of energy and will not grow
Yeast will only grow when protein X binds to protein Y to drive transcription
Confirming Protein/Protein binding - IP
Do these two proteins interact? YES - GST-FoxD3 and Oct-4 form a band - presence indicates that
it is interacting
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