TY - JOUR
T1 - Interaction of human DNA polymerase η with monoubiquitinated PCNA
T2 - A possible mechanism for the polymerase switch in response to DNA damage
AU - Kannouche, Patricia L.
AU - Wing, Jonathan
AU - Lehmann, Alan R.
N1 - Funding Information:
We are grateful to Jen Lees for assistance with DNA sequencing and generation of antibodies; M. Shivji for the pET39-hPCNA plasmid; Fumio Hanaoka for the CPD-containing template; Maria Fousteri and Barry Coull for helpful advice; and Tony Carr and Catherine Green for comments on the manuscript. This work was supported by an MRC program grant and EU FP5 QoL grant QLG-CT-1999-00181.
PY - 2004/5/21
Y1 - 2004/5/21
N2 - Most types of DNA damage block replication fork progression during DNA synthesis because replicative DNA polymerases are unable to accommodate altered DNA bases in their active sites. To overcome this block, eukaryotic cells employ specialized translesion synthesis (TLS) polymerases, which can insert nucleotides opposite damaged bases. In particular, TLS by DNA polymerase η (polη) is the major pathway for bypassing UV photoproducts. How the cell switches from replicative to TLS polymerase at the site of blocked forks is unknown. We show that, in human cells, PCNA becomes monoubiquitinated following UV irradiation of the cells and that this is dependent on the hRad18 protein. Monoubiquitinated PCNA but not unmodified PCNA specifically interacts with polη, and we have identified two motifs in polη that are involved in this interaction. Our findings provide an attractive mechanism by which monoubiquitination of PCNA might mediate the polymerase switch.
AB - Most types of DNA damage block replication fork progression during DNA synthesis because replicative DNA polymerases are unable to accommodate altered DNA bases in their active sites. To overcome this block, eukaryotic cells employ specialized translesion synthesis (TLS) polymerases, which can insert nucleotides opposite damaged bases. In particular, TLS by DNA polymerase η (polη) is the major pathway for bypassing UV photoproducts. How the cell switches from replicative to TLS polymerase at the site of blocked forks is unknown. We show that, in human cells, PCNA becomes monoubiquitinated following UV irradiation of the cells and that this is dependent on the hRad18 protein. Monoubiquitinated PCNA but not unmodified PCNA specifically interacts with polη, and we have identified two motifs in polη that are involved in this interaction. Our findings provide an attractive mechanism by which monoubiquitination of PCNA might mediate the polymerase switch.
UR - http://www.scopus.com/inward/record.url?scp=2442417331&partnerID=8YFLogxK
U2 - 10.1016/S1097-2765(04)00259-X
DO - 10.1016/S1097-2765(04)00259-X
M3 - Article
C2 - 15149598
AN - SCOPUS:2442417331
SN - 1097-2765
VL - 14
SP - 491
EP - 500
JO - Molecular Cell
JF - Molecular Cell
IS - 4
ER -