Huang et al' Notes

1
Huang et al. Notes
transcription factor protein that binds to
promoters or enhancers in the nucleus where
they are able to regulate transcription one key
regulator of transcription factors is the ability
to access the DNA transcription factors in the
cytoplasm are inactive NFkB activates many
genes, and is upregulated in prostate cancers
shown to regulate IL-8, MMP-9, and VEGF
normally inhibited by IkB which keeps it in the
cytoplasm IkB is degraded by ubiquitin
mediated degradation IkBaM is a mutated IkB
protein that cannot be degraded so always
acts ubiquitin small protein that marks
proteins for degradation by the proteasome
(ribosome-sized protein machine that degrades
proteins) IL-8 interleukin-8 growth factor for
several kinds of cells MMP-9 matrix
metalloprotease-9 breaks down extracellular
matrix (aids in invasiveness of tumors- gives
more access to cells and blood vessels
2
Huang et al. Notes
EMSA electro-mobility shift assay also referred
to as a gel shift assay technique used to
detect protein binding to short pieces of
DNA short pieces of DNA run through a gel
relatively fast toward the pole proteins
bound to the DNA makes the complex move more
slowly referred to as a gel shift
supershift further slowing of a DNAprotein
complex using antibodies that bind to the
protein that is bound to the DNA (larger
still) luciferase reporter enzyme that is
relatively easy to quantify amount of enzyme
directly related to how active the promoter is
originally found in fireflies-- gives off a
yellowish light luciferase from different
species have different requirements to be active
allows 2 different promoter activities to be
measured in the same cells
3
Huang et al. Notes
zymography method for determining the amount of
enzyme present uses an SDS gel with some kind
of matrix proteins in addition to
acrylamide to produce white spots on a black
background matrigel common support matrix for
tissue culture cells- contains lots of
collagen to support cells growing in a suspension