scholarly journals Ubiquitinated proliferating cell nuclear antigen activates translesion DNA polymerases   and REV1

2005 ◽  
Vol 102 (51) ◽  
pp. 18361-18366 ◽  
Author(s):  
P. Garg ◽  
P. M. Burgers
Keyword(s):  
Proliferating Cell Nuclear Antigen ◽  
Dna Polymerases ◽  
Nuclear Antigen ◽  
Translesion Dna ◽  
Proliferating Cell

scholarly journals Effect of Proliferating Cell Nuclear Antigen Ubiquitination and Chromatin Structure on the Dynamic Properties of the Y-family DNA Polymerases

2008 ◽  
Vol 19 (12) ◽  
pp. 5193-5202 ◽  
Author(s):  
Simone Sabbioneda ◽  
Audrey M. Gourdin ◽  
Catherine M. Green ◽  
Angelika Zotter ◽  
Giuseppina Giglia-Mari ◽  
...  
Keyword(s):  
Chromatin Structure ◽  
Proliferating Cell Nuclear Antigen ◽  
Dynamic Properties ◽  
Human Fibroblasts ◽  
Dna Polymerases ◽  
S Phase ◽  
Nuclear Antigen ◽  
Replication Foci ◽  
Y Family ◽  
Proliferating Cell

Y-family DNA polymerases carry out translesion synthesis past damaged DNA. DNA polymerases (pol) η and ι are usually uniformly distributed through the nucleus but accumulate in replication foci during S phase. DNA-damaging treatments result in an increase in S phase cells containing polymerase foci. Using photobleaching techniques, we show that polη is highly mobile in human fibroblasts. Even when localized in replication foci, it is only transiently immobilized. Although ubiquitination of proliferating cell nuclear antigen (PCNA) is not required for the localization of polη in foci, it results in an increased residence time in foci. polι is even more mobile than polη, both when uniformly distributed and when localized in foci. Kinetic modeling suggests that both polη and polι diffuse through the cell but that they are transiently immobilized for ∼150 ms, with a larger proportion of polη than polι immobilized at any time. Treatment of cells with DRAQ5, which results in temporary opening of the chromatin structure, causes a dramatic immobilization of polη but not polι. Our data are consistent with a model in which the polymerases are transiently probing the DNA/chromatin. When DNA is exposed at replication forks, the polymerase residence times increase, and this is further facilitated by the ubiquitination of PCNA.

scholarly journals DNA Polymerases BI and D from the Hyperthermophilic Archaeon Pyrococcus furiosus Both Bind to Proliferating Cell Nuclear Antigen with Their C-Terminal PIP-Box Motifs

2007 ◽  
Vol 189 (15) ◽  
pp. 5652-5657 ◽  
Author(s):  
Kazuo Tori ◽  
Megumi Kimizu ◽  
Sonoko Ishino ◽  
Yoshizumi Ishino
Keyword(s):  
Dna Synthesis ◽  
Proliferating Cell Nuclear Antigen ◽  
Pyrococcus Furiosus ◽  
Dna Polymerases ◽  
Nuclear Antigen ◽  
Hyperthermophilic Archaeon ◽  
Sliding Clamp ◽  
Replication Fork Progression ◽  
Proliferating Cell

ABSTRACT Proliferating cell nuclear antigen (PCNA) is the sliding clamp that is essential for the high processivity of DNA synthesis during DNA replication. Pyrococcus furiosus, a hyperthermophilic archaeon, has at least two DNA polymerases, polymerase BI (PolBI) and PolD. Both of the two DNA polymerases interact with the archaeal P. furiosus PCNA (PfuPCNA) and perform processive DNA synthesis in vitro. This phenomenon, in addition to the fact that both enzymes display 3′-5′ exonuclease activity, suggests that both DNA polymerases work in replication fork progression. We demonstrated here that both PolBI and PolD functionally interact with PfuPCNA at their C-terminal PIP boxes. The mutant PolBI and PolD enzymes lacking the PIP-box sequence do not respond to the PfuPCNA at all in an in vitro primer extension reaction. This is the first experimental evidence that the PIP-box motif, located at the C termini of the archaeal DNA polymerases, is actually critical for PCNA binding to form a processive DNA-synthesizing complex.

scholarly journals Structure of a Mutant Form of Proliferating Cell Nuclear Antigen That Blocks Translesion DNA Synthesis†‡

Biochemistry ◽  
2008 ◽  
Vol 47 (50) ◽  
pp. 13354-13361 ◽  
Author(s):  
Bret D. Freudenthal ◽  
S. Ramaswamy ◽  
Manju M. Hingorani ◽  
M. Todd Washington
Keyword(s):  
Dna Synthesis ◽  
Proliferating Cell Nuclear Antigen ◽  
Nuclear Antigen ◽  
Mutant Form ◽  
Translesion Dna Synthesis ◽  
Translesion Dna ◽  
Proliferating Cell

scholarly journals Somatic hypermutation of immunoglobulin genes: lessons from proliferating cell nuclear antigen K164R mutant mice

2008 ◽  
Vol 364 (1517) ◽  
pp. 621-629 ◽  
Author(s):  
Petra Langerak ◽  
Peter H.L Krijger ◽  
Marinus R Heideman ◽  
Paul C.M van den Berk ◽  
Heinz Jacobs
Keyword(s):  
B Cells ◽  
Proliferating Cell Nuclear Antigen ◽  
Somatic Mutations ◽  
Dna Polymerases ◽  
Critical Role ◽  
Dna Lesions ◽  
Nuclear Antigen ◽  
Immunoglobulin Genes ◽  
Proliferating Cell

Proliferating cell nuclear antigen (PCNA) encircles DNA as a ring-shaped homotrimer and, by tethering DNA polymerases to their template, PCNA serves as a critical replication factor. In contrast to high-fidelity DNA polymerases, the activation of low-fidelity translesion synthesis (TLS) DNA polymerases seems to require damage-inducible monoubiquitylation (Ub) of PCNA at lysine residue 164 (PCNA-Ub). TLS polymerases can tolerate DNA damage, i.e. they can replicate across DNA lesions. The lack of proofreading activity, however, renders TLS highly mutagenic. The advantage is that B cells use mutagenic TLS to introduce somatic mutations in immunoglobulin (Ig) genes to generate high-affinity antibodies. Given the critical role of PCNA-Ub in activating TLS and the role of TLS in establishing somatic mutations in immunoglobulin genes, we analysed the mutation spectrum of somatically mutated immunoglobulin genes in B cells from PCNA K164R knock-in mice. A 10-fold reduction in A/T mutations is associated with a compensatory increase in G/C mutations—a phenotype similar to Polη and mismatch repair-deficient B cells. Mismatch recognition, PCNA-Ub and Polη probably act within one pathway to establish the majority of mutations at template A/T. Equally relevant, the G/C mutator(s) seems largely independent of PCNA K164 modification.

scholarly journals Functional Interactions of a Homolog of Proliferating Cell Nuclear Antigen with DNA Polymerases inArchaea

1999 ◽  
Vol 181 (21) ◽  
pp. 6591-6599 ◽  
Author(s):  
Isaac K. O. Cann ◽  
Sonoko Ishino ◽  
Ikuko Hayashi ◽  
Kayoko Komori ◽  
Hiroyuki Toh ◽  
...  
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Proliferating Cell Nuclear Antigen ◽  
Dna Polymerases ◽  
Nuclear Antigen ◽  
Pol Ii ◽  
Circular Dna ◽  
Pol I ◽  
Factor C ◽  
Proliferating Cell

ABSTRACT Proliferating cell nuclear antigen (PCNA) is an essential component of the DNA replication and repair machinery in the domainEucarya. We cloned the gene encoding a PCNA homolog (PfuPCNA) from an euryarchaeote, Pyrococcus furiosus, expressed it in Escherichia coli, and characterized the biochemical properties of the gene product. The protein PfuPCNA stimulated the in vitro primer extension abilities of polymerase (Pol) I and Pol II, which are the two DNA polymerases identified in this organism to date. An immunological experiment showed that PfuPCNA interacts with both Pol I and Pol II. Pol I is a single polypeptide with a sequence similar to that of family B (α-like) DNA polymerases, while Pol II is a heterodimer. PfuPCNA interacted with DP2, the catalytic subunit of the heterodimeric complex. These results strongly support the idea that the PCNA homolog works as a sliding clamp of DNA polymerases in P. furiosus, and the basic mechanism for the processive DNA synthesis is conserved in the domainsBacteria, Eucarya, and Archaea. The stimulatory effect of PfuPCNA on the DNA synthesis was observed by using a circular DNA template without the clamp loader (replication factor C [RFC]) in both Pol I and Pol II reactions in contrast to the case of eukaryotic organisms, which are known to require the RFC to open the ring structure of PCNA prior to loading onto a circular DNA. Because RFC homologs have been found in the archaeal genomes, they may permit more efficient stimulation of DNA synthesis by archaeal DNA polymerases in the presence of PCNA. This is the first stage in elucidating the archaeal DNA replication mechanism.

Proliferating cell nuclear antigen: More than a clamp for DNA polymerases

BioEssays ◽  
1997 ◽  
Vol 19 (11) ◽  
pp. 967-975 ◽  
Author(s):  
Zophonías O. Jónsson ◽  
Ulrich Hübscher
Keyword(s):  
Proliferating Cell Nuclear Antigen ◽  
Dna Polymerases ◽  
Nuclear Antigen ◽  
Proliferating Cell

scholarly journals The Post-translational Modifications of Proliferating Cell Nuclear Antigen

2004 ◽  
Vol 279 (19) ◽  
pp. 20194-20199 ◽  
Author(s):  
Stanislav N. Naryzhny ◽  
Hoyun Lee
Keyword(s):  
Dna Replication ◽  
Proliferating Cell Nuclear Antigen ◽  
Dna Polymerases ◽  
Nuclear Antigen ◽  
Two Dimensional ◽  
Basic Form ◽  
Post Translational Modifications ◽  
Acetylation Status ◽  
Acidic Form ◽  
Proliferating Cell

The diverse function of proliferating cell nuclear antigen (PCNA) is thought to be due, in large part, to post-translational modifications. Here we show by high resolution two-dimensional PAGE analysis that there are three distinct PCNA isoforms that differ in their acetylation status. The moderately acetylated main (M) form was found in all of the subcellular compartments of cycling cells, whereas the highly acetylated acidic form was primarily found in the nucleoplasm, nuclear matrix, and chromatin. Interestingly, the deacetylated basic form was most pronounced in the nucleoplasm of cycling cells. The cells in G0and the cytoplasm of cycling cells contained primarily the M form only. Because p300 and histone deacetylase (HDAC1) were co-immunoprecipitated with PCNA, they are likely responsible for the acetylation and deacetylation of PCNA, respectively. We also found that deacetylation reduced the ability of PCNA to bind to DNA polymerases β and δ. Taken together, our data support a model where the acidic and M forms participate in DNA replication, whereas the basic form is associated with the termination of DNA replication.

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