rest regulatory networks
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REST regulatory networks
Gene Yeo1,2 Stefan Aigner 2, Eric Van Nostrand1,2, Fred H. Gage2
1Crick-Jacobs Center of Theoretical and Computational Biology 2Laboratory of Genetics
The Salk Institute for Biological Studies, La Jolla, CA
REST repressor/silencing complex silences neuronal genes in non-neuronal
cells
Differentiated non-neuronal cells
REST
mSin3MeCP2 HDAC SCP
CoREST
HDAC
MeCP2
Histone K4 Demethylase
HMT K9
HP1
DNMT1
RE1mK9
mK9
mK9
mK9 mK9
mK9
mK9
mK9
m
m m m m
m
mm m
m mCpG
mK9 Methyl H3K9
Slide courtesy of XC
REST binds to a ~20 bp sequenceREST levels decrease during neuronal differentiation
REST sites are highly-conserved across evolution
Brain-derived neurotrophic factor
Superiorcervical ganglion neural-specific 10
Synaptosomal-associated protein 25
Algorithm to identify conserved REST sites
Converged motif
Several REST sites are proximal to microRNAs
REST-regulated microRNAs are neuronal
REST binds in vivo to predicted sites
REST occupancy near microRNA genes decreases during neural differentiation
Anti-REST chromatin immunoprecipitation/qPCR for candidate REST sites in P19 cells upon neural induction
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
miR-9-3
miR-29a/29b-1
30e/30c-1 miR135b miR153-1
miR182/96/183miR192/194-2miR219-1
% of chromatin immunoprecipitated
Undifferentiated
Differentiated
Neural-specific microRNAs and REST co-repressors during mouse stem cell
neuronal differentiation
d7d5 d9 d11d0
Neuronal marker TUJ1
REST
msin3A
SCP-1
CoREST
Neural differentiation of P19 mouse embryonic carcinoma cells
d7d5 d9 d11d0
miR-9
miR-124a
U6
Northern blots Western blots
RA induction
Neural-specific microRNAs likely target com-
ponents of the REST co-repressor complex
Plasmid transfected: GFP empty mir-9
-actin
mir1
24
mir1
mir3
24
empty
msin3A SCP-1
Loadingcontrol
Plasmid transfected:
mir-9 and mir-124a downregulate mSin3a and SCP-1, respectively
REST
mSin3MeCP2 HDAC SCP
CoREST
HDAC
MeCP2
Histone K4 Demethylase
HMT K9
HP1
DNMT1
RE1mK9
mK9
mK9
mK9 mK9
mK9
mK9
mK9
m
m m m m
m
mm m
m mCpG
mK9 Methyl H3K9
Neural-specific microRNAs likely target com-
ponents of the REST corepressor complex A sensor assay confirms predicted target sites for miR-124a in the
3’UTR of SCP-1
SCP-1 sensor
Control sensor
SCP-1 3’UTR
ADAR1 3’UTR
Control sensor SCP1 sensor + mir124
Enhancing neurogenesis mir-124 induces neuronal gene expression
in P19 mouse EC cells
0
1
2
3
4
5
6
GAP43 NFH Glur2 ND1
Promoter driving luciferase
Fold change
(mir124/control)
24 hours
48 hours
REST high
Neuronal progenitors Neurons
REST-regulated microRNAs OFF
REST-regulated microRNAs ON
REST-regulated microRNAs downregulate components of the REST repressor complex
REST-regulated protein coding genes ON
REST-regulated protein coding genes OFF
A model for REST regulation to enhance neurogenesis
REST low
Acknowledgements: Eunice Meija, Xinwei Cao Supported by Crick-Jacobs Center, NSF
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