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Footprinting DNA-Protein Interactions
Footprinting DNA-Protein Interactions
Interactions
• Powerful and fairly rapid methods for
mapping where and how proteins bind
tightly to DNA
• 2 ways:
1. DNAse I footprinting
2. DMS footprinting
DNAse I Footprinting
Fig. 5.37a
Fig. 5.37b
Sample of a DNase I
footprinting gel.
Footprint
Fig. 5.38a
Example of DMS footprinting.
Fig. 5.38b
Positive Control of Lac Operon
• Catabolite Repression hypothesis
– predicted that glucose would inhibit synthesis of
other sugar metabolizing pathway enzymes (e.g.,
lactose pathway)
• Partially right, its lack of activation instead of true
repression
– Cells respond to high glucose with lowered levels
of cAMP and vice-versa
– cAMP activates Lac operon via CAP
cyclic 5’-3’ phosphodiester in cAMP
glucose cAMP
• CAP-cAMP increases Kb
Lac Control Region
Fig. 7.16
CAP-cAMP dimer interacts with the CTD of the
a subunits of the RNAP Core
Fig 7.17
Why does the Lac Operon need an activator?
-35 -10
TTTACAC ---------------- TATGTT (Lac)
-35 -10
TTGACAT --------------- TATAAT (consensus)
Fig. 7.31
Fig. 7.32
Biological advantage: