PDBsum entry 1jsp

DNA binding protein

PDB id: 1jsp

Contents

Protein chains

20 a.a.

121 a.a. *

* Residue conservation analysis

PDB id:

1jsp

Name:

DNA binding protein

Title:

Nmr structure of cbp bromodomain in complex with p53 peptide

Structure:

Tumor protein p53. Chain: a. Fragment: c-terminal fragment. Engineered: yes. Creb-binding protein. Chain: b. Fragment: bromodomain. Synonym: cbp. Engineered: yes

Source:

Synthetic: yes. Other_details: this sequence occurs naturally in humans.. Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli bl21. Expression_system_taxid: 511693.

NMR struc:

20 models

Authors:

Y.He,S.Mujtaba,L.Zeng,S.Yan,M.-M.Zhou

Key ref:

S.Mujtaba et al. (2004). Structural mechanism of the bromodomain of the coactivator CBP in p53 transcriptional activation. Mol Cell, 13, 251-263. PubMed id: 14759370 DOI: 10.1016/S1097-2765(03)00528-8

Date:

17-Aug-01 Release date: 17-Aug-02

Protein chain

P04637  (P53_HUMAN) -  Cellular tumor antigen p53 from Homo sapiens

Seq: 393 a.a.

Struc: 20 a.a.*

Protein chain

Q92793  (CBP_HUMAN) -  CREB-binding protein from Homo sapiens

Seq: 2442 a.a.

Struc: 121 a.a.*

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

Enzyme reactions

• Enzyme class 1: Chain B: E.C.2.3.1.- - ?????
• Enzyme class 2: Chain B: E.C.2.3.1.48 - histone acetyltransferase.
• Reaction: L-lysyl-[protein] + acetyl-CoA = N₆-acetyl-L-lysyl-[protein] + CoA + H⁺

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

DOI no: 10.1016/S1097-2765(03)00528-8

Mol Cell 13:251-263 (2004)

PubMed id: 14759370

Structural mechanism of the bromodomain of the coactivator CBP in p53 transcriptional activation.

S.Mujtaba, Y.He, L.Zeng, S.Yan, O.Plotnikova, Sachchidanand, R.Sanchez, N.J.Zeleznik-Le, Z.Ronai, M.M.Zhou.

ABSTRACT

Lysine acetylation of the tumor suppressor protein p53 in response to a wide variety of cellular stress signals is required for its activation as a transcription factor that regulates cell cycle arrest, senescence, or apoptosis. Here, we report that the conserved bromo-domain of the transcriptional coactivator CBP (CREB binding protein) binds specifically to p53 at the C-terminal acetylated lysine 382. This bromodomain/acetyl-lysine binding is responsible for p53 acetylation-dependent coactivator recruitment after DNA damage, a step essential for p53-induced transcriptional activation of the cyclin-dependent kinase inhibitor p21 in G1 cell cycle arrest. We further present the three-dimensional nuclear magnetic resonance structure of the CBP bromodomain in complex with a lysine 382-acetylated p53 peptide. Using structural and biochemical analyses, we define the molecular determinants for the specificity of this molecular recognition.

Selected figure(s)

Figure 4.
Figure 4. Functional Role of the CBP Bromodomain in CBP/p53 Association(A) Effect of CBP bromodomain on p53-induced p21 activation in 10.1 cells after UV treatment. p21 activity of the 10.1 cells transfected with p53 or p53 mutant together with p21 luciferase and β-galactosidase, with or without CBP bromodomain, was measured in a luciferase-based assay. Mean values of the luciferase activities represent at least three independent cell transfections.(B) Western blotting analysis assessing protein expression in the transfected 10.1 cells. Note that numerals below the p21 blot represent ratio of p21 expression in the cells transfected with p53, with or without CBP bromodomain, to that in the cells transfected with only the empty vector pCDNA3. The signals were quantitated using Kodak 1D Digital Image Analysis Software.(C) Assessing effects of cotransfected bromodomains on p53-induced p21 activation in the 10.1 cells. The cell transfections and p21 luciferase activity analysis were same as described in (A).(D) Western blots assessing expression of various bromodomains and p53 in the transfected 10.1 cells with or without UV-C treatment.(E) Effect of the CBP bromodomain on cell cycle distribution induced by p53 in the 10.1 cells transfected with Us9-GFP, wild-type, or mutant p53, with or without the CBP bromodomains. The DNA content of the gated GFP-positive cells in G1 phase was determined by PI staining and FACS analysis. Average values of the DNA content of the cell cycle phases in each different experiment represent at least three independent transfection trials.

Figure 6.
Figure 6. Mutational Analyses of CBP Bromodomain Binding to p53(A) Effect of point mutation of CBP bromodomain residues on p53 peptide binding. Western blot with GST antibody shows GST-CBP bromodomain binding to the biotinylated p53 AcK382 peptide bound to streptavidin-agarose beads (top). Relatively equal amount of proteins was used in each binding experiment (bottom). Mutants highlighted in red exhibited markedly reduced binding to the p53 peptide. Mutational effects on the protein/peptide binding were quantitated.(B) Mutational analysis of p53 peptide residues, assessed in a competition assay as described in Experimental Procedures. Mutant p53 peptides that showed a major reduction in binding to the bromodomain are indicated in red.

The above figures are reprinted by permission from Cell Press: Mol Cell (2004, 13, 251-263) copyright 2004.

Figures were selected by an automated process.