A p53 enhancer region regulates target genes through

chromatin conformations in cis and in trans.

Nichole Link, Paula Kurtz, Melissa ONeal,
Gianella Garcia-Hughes, and John M. Abrams

Presented by Aleksandra, Ambra, Lyudmil, Manuel.

PLAN
1. INTRODUCTION
2. RESULTS & METHODS
3. CONCLUSIONS

INTRODUCTION

INTRODUCTION

Figure 1 A (fragment) .
Oncogene. 2007 Aug 9; 26(36):5184-93.

INTRODUCTION

Figure 1 A (fragment) .
Oncogene. 2007 Aug 9; 26(36):5184-93.

RESULTS

FRT-mediated recombination

Fig 1 A, B.
Nature Protocols 6, 14121428 (2011)

Deletions

Fig 1 A
Genes Dev. Nov 15, 2013; 27(22): 24332438.

RESULTS

p53-dependent stress-induced cell death

Acridine stains

Fig 1 E, F.
Genes Dev. Nov 15, 2013; 27(22): 24332438.

RESULTS

P53 is expressed in
the D2p53RE mutant

RIPD and control cDNAs

Fig 1 B, C, D.
Supplementary 1 A, B.
Genes Dev. Nov 15, 2013; 27(22): 24332438.

Revisting 3C(Chromosome Conformation

Capture)
Is this due to the formation of chromosome loops?

 Normalized to known interactions in bitorax complex

No p53

Irradiation

 Is the p53RE present ectopically able to establish this contacts? Include

it in other chromosome(2)

FISH experiment

Fig.2
p53RE stablishes contact with its regulatory regions independently of:

Stress situation
Pressence of the p53 protein
Therefore, this contacts do not induct transcription by themselves.
This contacts can be rescued by ectopic presence of the p53RE.

p53RE is capable of stablishing contacts in trans, from far

regions.

RESULTS
Long distance control by the p53RE

xrp1 is nonresponsive in D2p53RE animals and D3control

stress-induced expression unchanged in D3control

RESULTS

xrp1
p53RE

p53RE regulates xrp1

colocalized
signal

RESULTS

In vivo frequency of contact between

p53 enhancer and xrp1

p53 enhancer makes physical contact to trans targets within only a subset of
cells at any given time

RESULTS

p53RE can perform contacts with long-distance target sites from ectopic position in
trans in a sequence-specific way.

RESULTS

p53RE partially restores regulation from an ectopic position in trans

Presence of contacts not sufficient for target

RESULTS
p53RE can generate simultaneous contacts with multiple targets in a single cell

colocalization of al three loci within 23% of cells

CONCLUSIONS
Single enhancer region

regulation of multiple targets in cis and in trans

Stimulus dependent
induction of multiple genes
4 kb to 330 kb

p53
regulatory
element

xrp1
a genetically unlinked target residing
across the centromere

Transplanted to ectopic locations

contacts with target sites reestablished
regulation of some target genes was restored

long-range regulation by the p53RE involves chromosomal architectures that link this
enhancer to target genes regardless of whether they are in cis or in trans.