TBP, TATA binding protein;

Ai miei genitori , Sandro e Maria

List of Abbreviations.

TFIID, transcription factor II D;

TBP, TATA binding protein;

TAFs, TBP associated factor;

TFIIH, transcription factor II H;

TATA, sequence of thymine-adenine-thymine-adenine;

MDM-2, ubiquitin E

ligase Murine Double Minute 2 protein;

ATM kinase, ataxia telangectasia mutated;

CDK, cycline-dependent kinase;

MIZ1, MYC interacting zinc-finger protein-1;

SP1, specificity protein 1;

ZAC1, zinc-finger protein;

ZBTB4, zinc-finger and BTB domain containing 4;

MYC, transcription factor;

BTB, broad-complex , tramtrack and bric-abrac;

POZ, poxvirus and zinc finger;

HuR (ELAVL1), human antigen R ( ELAV-like protein I );

ARE, AU-rich element;

UTR, untranslated region;

RNPC1 or RBM38, RNA binding motif 38;

CUGBP1, CUG triplet repeat RNA binding protein 1;

CALR, calreticulin;

AKT, protein kinase B ( PKB );

PCNA, proliferating cell nuclear antigen;

CDK4, cicli dependent kinase 4;

PIM1, proto-oncogene serine/threonine-protein kinase;

P38α, mitogen-activated protein kinase;

JNK1, protein kinase of the MAPK family;

GSK3β, glycogen synthase kinase 3 beta;

SCF, Skp , Cullin , F-box containing complex;

Skp2, S-phase kinase-associated protein 2;

SCFSkp2, ubiquitin ligase;

APC, anaphase promoting complex;

Cdc20, cell-division cycle protein 20;

CRL4Cdt2, E

ubiquitin ligase complex promotes PCNA monoubiquitination in proliferating cells;

Hsp90, heat shock protein;

BRCA1, human caretaker gene that produce a protein called breast cancer type 1;

SUV39H1, histone-lysine N-methyltransferase;

DNMT3A, DNA (cytosine-5)-methyltransferase 3A;

Cd95, cluster of differentiation;

Apo1, apoptosis antigen 1;

TNF-R, tumor necrosis factor receptor;

FADD, Fas-Associated protein with Death Domain;

DR, death receptor;

PERP, pro-apoptotic gene;

TRAIL, TNF-related apoptosis-inducing ligand;

PTEN, phosphatase and tensin homolog;

PIP2, phosphatidyl-inositol-2-phosphate;

PI3K, phosphatidylinositol 3-kinase;

MOMP, mithocondrial outermembrane permeabilization;

Bax, Bcl-2-associated X;

Bak, Bcl-2 antagonist/killer;

Apaf, apoptotic peptidase activating factor 1;

BH, Bcl-2 homology;

PUMA, p53 upregulated modulator of apoptosis;

ASC, apoptosis-associated spekle-like protein;

tBid, truncated Bid;

Rb, retinoblastoma;

PP2A, protein phosphatise 2A;

CREB1, CAMP responsive element binding protein 1;

CBP, CREB-binding-protein;

SCID, severe combined immune deficiency.

Index

Intoduction 1

P53: structure and functions 3

Stabilization of p53 in response to DNA damage 6

P53 pathway 7

The p21 circuit 8

Target protein 14-3-3 10

The extrinsic apoptotic pathway 11

The intrinsic apoptotic pathway 12

Positive and negative p53 feedback loops 16

P53-MDM2 interaction 21

Structural requirements for p53-MDM2 interaction p21 22

P53 as a therapeutic target 24

Blocking MDM2 exprection 25

Inhibiting MDM2 ubiquitin ligase activity 26

Inhibiting MDM2-p53 binding 26

Small-molecule inhibitors of the p53-MDM2 interaction 27

Introduction to experimental section 30

Experimental section 42

Materials and Methods 43

Chemistry 44

References 50