Targeting of Human DNA Polymerase ι to the Replication Machinery via Interaction with PCNA

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 replica...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 98; no. 25; pp. 14256 - 14261
Main Authors Haracska, Lajos, Johnson, Robert E., Unk, Ildiko, Phillips, Barbara B., Hurwitz, Jerard, Prakash, Louise, Prakash, Satya
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 04.12.2001
National Acad Sciences
The National Academy of Sciences
Subjects
Base Sequence
Binding Sites
Biochemistry
Biological Sciences
Cells
Deoxyribonucleic acid
Dimers
DNA
DNA - biosynthesis
DNA - genetics
DNA Damage
DNA Polymerase iota
DNA Primers - genetics
DNA Replication - physiology
DNA-Binding Proteins - metabolism
DNA-Directed DNA Polymerase - metabolism
Gels
Humans
In Vitro Techniques
Kinetics
Lesions
Molecular Sequence Data
Nucleotides
Oligodeoxyribonucleotides - metabolism
Oligomers
pol^i protein
Polls
Proliferating Cell Nuclear Antigen - metabolism
replication factor C
Replication Protein A
Replication Protein C
Studies
Yeasts
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Summary: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 hPolι is increased ≈80-150-fold, and this increase in efficiency results from a reduction in the apparent Kmfor 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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To whom reprint requests should be addressed. E-mail: sprakash@scms.utmb.edu.
Contributed by Jerard Hurwitz
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.261560798