PDBsum entry 3l0w

Replication

PDB id: 3l0w

Contents

Protein chains

163 a.a. *

166 a.a. *

* Residue conservation analysis

PDB id:

3l0w

Name:

Replication

Title:

Structure of split monoubiquitinated pcna with ubiquitin in position two

Structure:

Proliferating cell nuclear antigen. Chain: a. Fragment: n fragment. Synonym: pcna. Engineered: yes. Monoubiquitinated proliferating cell nuclear antigen. Chain: b. Fragment: ubi-c fragment. Engineered: yes

Source:

Saccharomyces cerevisiae. Brewer's yeast,lager beer yeast,yeast. Organism_taxid: 4932. Gene: pol30, ybr0811, ybr088c. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: pol30, ubi1 & pol30, ybr0811, ybr088c.

Resolution:

2.80Å R-factor: 0.279 R-free: 0.314

Authors:

B.D.Freudenthal,L.Gakhar,S.Ramaswamy,M.T.Washington

Key ref:

B.D.Freudenthal et al. (2010). Structure of monoubiquitinated PCNA and implications for translesion synthesis and DNA polymerase exchange. Nat Struct Biol, 17, 479-484. PubMed id: 20305653

Date:

10-Dec-09 Release date: 23-Mar-10

Protein chain

P15873  (PCNA_YEAST) -  Proliferating cell nuclear antigen from Saccharomyces cerevisiae (strain ATCC 204508 / S288c)

Seq: 258 a.a.

Struc: 163 a.a.

Protein chain

P05759  (RS31_YEAST) -  Ubiquitin-ribosomal protein eS31 fusion protein from Saccharomyces cerevisiae (strain ATCC 204508 / S288c)

Seq: 152 a.a.

Struc: 166 a.a.*

Protein chain

P15873  (PCNA_YEAST) -  Proliferating cell nuclear antigen from Saccharomyces cerevisiae (strain ATCC 204508 / S288c)

Seq: 258 a.a.

Struc: 166 a.a.*

* PDB and UniProt seqs differ at 145 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chains A, B: E.C.?

Nat Struct Biol 17:479-484 (2010)

PubMed id: 20305653

Structure of monoubiquitinated PCNA and implications for translesion synthesis and DNA polymerase exchange.

B.D.Freudenthal, L.Gakhar, S.Ramaswamy, M.T.Washington.

ABSTRACT

DNA synthesis by classical polymerases can be blocked by many lesions. These blocks are overcome by translesion synthesis, whereby the stalled classical, replicative polymerase is replaced by a nonclassical polymerase. In eukaryotes this polymerase exchange requires proliferating cell nuclear antigen (PCNA) monoubiquitination. To better understand the polymerase exchange, we developed a means of producing monoubiquitinated PCNA, by splitting the protein into two self-assembling polypeptides. We determined the X-ray crystal structure of monoubiquitinated PCNA and found that the ubiquitin moieties are located on the back face of PCNA and interact with it through their canonical hydrophobic surface. Moreover, the attachment of ubiquitin does not change PCNA's conformation. We propose that PCNA ubiquitination facilitates nonclassical polymerase recruitment to the back of PCNA by forming a new binding surface for nonclassical polymerases, consistent with a 'tool belt' model of the polymerase exchange.