Abstract
Treatment of yeast and human cells with DNA-damaging agents elicits Rad6-Rad18-mediated monoubiquitination of proliferating cell nuclear antigen (PCNA) at its Lys-164 residue [ubiquitin (Ub)-PCNA], and this PCNA modification is indispensable for promoting the access of translesion synthesis (TLS) polymerases (Pols) to PCNA. However, the means by which K164-linked Ub modulates the proficiency of TLS Pols to bind PCNA and take over synthesis from the replicative Pol has remained unclear. One model that has gained considerable credence is that the TLS Pols bind PCNA at 2 sites, to the interdomain connector loop via their PCNA-interacting protein (PIP) domain and to the K164-linked Ub moiety via their Ub-binding domain (UBD). Specifically, this model postulates that the UBD-mediated binding of TLS Pols to the Ub moiety on PCNA is necessary for TLS. To test the validity of this model, we examine the contributions that the PIP and Ub-binding zinc finger (UBZ) domains of human Polη make to its functional interaction with PCNA, its colocalization with PCNA in replication foci, and its role in TLS in vivo. We conclude from these studies that the binding to PCNA via its PIP domain is a prerequisite for Polη's ability to function in TLS in human cells and that the direct binding of the Ub moiety on PCNA via its UBZ domain is not required. We discuss the possible role of the Ub moiety on PCNA in TLS.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 17724-17729 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 105 |
| Issue number | 46 |
| DOIs | |
| State | Published - Nov 18 2008 |
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Keywords
- Exchange
- PCNA ubiquitination
- PIP domain
- UBZ domain
ASJC Scopus subject areas
- General
Cite this
Roles of PCNA-binding and ubiquitin-binding domains in human DNA polymerase η in translesion DNA synthesis. / Acharya, Narottam; Yoon, Jung Hoon; Gali, Himabindu; Unk, Ildiko; Haracska, Lajos; Johnson, Robert E.; Hurwitz, Jerard; Prakash, Louise; Prakash, Satya.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, No. 46, 18.11.2008, p. 17724-17729.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Roles of PCNA-binding and ubiquitin-binding domains in human DNA polymerase η in translesion DNA synthesis
AU - Acharya, Narottam
AU - Yoon, Jung Hoon
AU - Gali, Himabindu
AU - Unk, Ildiko
AU - Haracska, Lajos
AU - Johnson, Robert E.
AU - Hurwitz, Jerard
AU - Prakash, Louise
AU - Prakash, Satya
PY - 2008/11/18
Y1 - 2008/11/18
N2 - Treatment of yeast and human cells with DNA-damaging agents elicits Rad6-Rad18-mediated monoubiquitination of proliferating cell nuclear antigen (PCNA) at its Lys-164 residue [ubiquitin (Ub)-PCNA], and this PCNA modification is indispensable for promoting the access of translesion synthesis (TLS) polymerases (Pols) to PCNA. However, the means by which K164-linked Ub modulates the proficiency of TLS Pols to bind PCNA and take over synthesis from the replicative Pol has remained unclear. One model that has gained considerable credence is that the TLS Pols bind PCNA at 2 sites, to the interdomain connector loop via their PCNA-interacting protein (PIP) domain and to the K164-linked Ub moiety via their Ub-binding domain (UBD). Specifically, this model postulates that the UBD-mediated binding of TLS Pols to the Ub moiety on PCNA is necessary for TLS. To test the validity of this model, we examine the contributions that the PIP and Ub-binding zinc finger (UBZ) domains of human Polη make to its functional interaction with PCNA, its colocalization with PCNA in replication foci, and its role in TLS in vivo. We conclude from these studies that the binding to PCNA via its PIP domain is a prerequisite for Polη's ability to function in TLS in human cells and that the direct binding of the Ub moiety on PCNA via its UBZ domain is not required. We discuss the possible role of the Ub moiety on PCNA in TLS.
AB - Treatment of yeast and human cells with DNA-damaging agents elicits Rad6-Rad18-mediated monoubiquitination of proliferating cell nuclear antigen (PCNA) at its Lys-164 residue [ubiquitin (Ub)-PCNA], and this PCNA modification is indispensable for promoting the access of translesion synthesis (TLS) polymerases (Pols) to PCNA. However, the means by which K164-linked Ub modulates the proficiency of TLS Pols to bind PCNA and take over synthesis from the replicative Pol has remained unclear. One model that has gained considerable credence is that the TLS Pols bind PCNA at 2 sites, to the interdomain connector loop via their PCNA-interacting protein (PIP) domain and to the K164-linked Ub moiety via their Ub-binding domain (UBD). Specifically, this model postulates that the UBD-mediated binding of TLS Pols to the Ub moiety on PCNA is necessary for TLS. To test the validity of this model, we examine the contributions that the PIP and Ub-binding zinc finger (UBZ) domains of human Polη make to its functional interaction with PCNA, its colocalization with PCNA in replication foci, and its role in TLS in vivo. We conclude from these studies that the binding to PCNA via its PIP domain is a prerequisite for Polη's ability to function in TLS in human cells and that the direct binding of the Ub moiety on PCNA via its UBZ domain is not required. We discuss the possible role of the Ub moiety on PCNA in TLS.
KW - Exchange
KW - PCNA ubiquitination
KW - PIP domain
KW - UBZ domain
UR - http://www.scopus.com/inward/record.url?scp=56649124781&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=56649124781&partnerID=8YFLogxK
U2 - 10.1073/pnas.0809844105
DO - 10.1073/pnas.0809844105
M3 - Article
VL - 105
SP - 17724
EP - 17729
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 46
ER -