TY - JOUR
T1 - Targeting of human DNA polymerase ι to the replication machinery via interaction with PCNA
AU - Haracska, Lajos
AU - Johnson, Robert E.
AU - Unk, Ildiko
AU - Phillips, Barbara B.
AU - Hurwitz, Jerard
AU - Prakash, Louise
AU - Prakash, Satya
PY - 2001/12/4
Y1 - 2001/12/4
N2 - Human DNA polymerase ι(hPolι) promotes translesion synthesis by inserting nucleotides opposite highly distorting or noninstructional DNA lesions. Here, we provide evidence for the physical interaction of hPolι with proliferating cell nuclear antigen (PCNA), and show that PCNA, together with replication factor C (RFC) and replication protein A (RPA), stimulates the DNA synthetic activity of hPolι. In the presence of these protein factors, on undamaged DNA, the efficiency (Vmax/Km) of correct nucleotide incorporation by hPoι is increased ≈80-150-fold, and this increase in efficiency results from a reduction in the apparent Km, for the nucleotide. PCNA, RFC, and RPA also stimulate nucleotide incorporation opposite the 3′-T of the (6-4) thymine-thymine (T-T) photoproduct and opposite an abasic site. The interaction of hPolι with PCNA implies that the targeting of this polymerase to the replication machinery stalled at a lesion site is achieved via this association.
AB - Human DNA polymerase ι(hPolι) promotes translesion synthesis by inserting nucleotides opposite highly distorting or noninstructional DNA lesions. Here, we provide evidence for the physical interaction of hPolι with proliferating cell nuclear antigen (PCNA), and show that PCNA, together with replication factor C (RFC) and replication protein A (RPA), stimulates the DNA synthetic activity of hPolι. In the presence of these protein factors, on undamaged DNA, the efficiency (Vmax/Km) of correct nucleotide incorporation by hPoι is increased ≈80-150-fold, and this increase in efficiency results from a reduction in the apparent Km, for the nucleotide. PCNA, RFC, and RPA also stimulate nucleotide incorporation opposite the 3′-T of the (6-4) thymine-thymine (T-T) photoproduct and opposite an abasic site. The interaction of hPolι with PCNA implies that the targeting of this polymerase to the replication machinery stalled at a lesion site is achieved via this association.
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U2 - 10.1073/pnas.261560798
DO - 10.1073/pnas.261560798
M3 - Article
C2 - 11724965
AN - SCOPUS:0035807870
SN - 0027-8424
VL - 98
SP - 14256
EP - 14261
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
IS - 25
ER -