The Non-canonical Protein Binding Site at the Monomer-Monomer Interface of Yeast Proliferating Cell Nuclear Antigen (PCNA) Regulates the Rev1-PCNA Interaction and Polζ/Rev1-dependent Translesion DNA Synthesis

In this study, experimental conditions are described that allowed us to follow the fate of the DNA polymerase delta-associated proliferating cell nuclear antigen (PCNA), by immunolabeling during the overall cell cycle. Differences in subcellular localization or the presence of PCNA allowed us to identify each phase of the cell cycle. Using these cell cycle markers in dog thyroid epithelial cells in primary culture, we found unexpected differences in cell cycle kinetics, in response to stimulations through cAMP-dependent and cAMP-independent pathways. These provide a new dimension to the view that the two pathways are largely separate, but co-operate on DNA synthesis initiation. More precisely, thyrotropin (TSH), acting via cAMP, exerts a potent triggering effect on DNA synthesis, associated with a precocious induction of PCNA appearance. This constitutes the major influence of TSH (cAMP) in determining cell cycle progression, which is only partly moderated by TSH-dependent lengthening of S- and G2-phases.

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The Influence of the Proliferating Cell Nuclear Antigen-interacting Domain of p21CIP1on DNA Synthesis Catalyzed by the Human andSaccharomyces cerevisiaePolymerase δ Holoenzymes

Journal of Biological Chemistry ◽

10.1074/jbc.272.4.2373 ◽

1997 ◽

Vol 272 (4) ◽

pp. 2373-2381 ◽

Cited By ~ 38

Author(s):

Emma Gibbs ◽

Zvi Kelman ◽

Jacqueline M. Gulbis ◽

Mike O'Donnell ◽

John Kuriyan ◽

...

Keyword(s):

Dna Synthesis ◽

Proliferating Cell Nuclear Antigen ◽

Nuclear Antigen ◽

Proliferating Cell ◽

Interacting Domain

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Stimulation of DNA synthesis by natural ceramide 1-phosphate

Biochemical Journal ◽

10.1042/bj3250435 ◽

1997 ◽

Vol 325 (2) ◽

pp. 435-440 ◽

Cited By ~ 75

Author(s):

Antonio GOMEZ-MUÑOZ ◽

Laura M. FRAGO ◽

Luis ALVAREZ ◽

Isabel VARELA-NIETO

Keyword(s):

Dna Synthesis ◽

Proliferating Cell Nuclear Antigen ◽

Solvent Mixture ◽

Mitogen Activated Protein Kinase ◽

Nuclear Antigen ◽

Tyrosine Residues ◽

Mitogen Activated Protein ◽

Ceramide 1 ◽

Proliferating Cell ◽

Stimulation Of

We found that natural (long-chain) ceramide 1-phosphate can be dispersed into aqueous solution when dissolved in an appropriate mixture of methanol/dodecane (49:1, v/v). This solvent mixture facilitates the interaction of this phosphosphingolipid with cells. Under these conditions, incubation of EGFR T17 fibroblasts with natural ceramide 1-phosphate caused a potent stimulation of DNA synthesis. This effect was accompanied by an increase in the levels of proliferating-cell nuclear antigen. Concentrations of natural ceramide 1-phosphate that stimulated the synthesis of DNA did not inhibit adenylate cyclase activity, nor did they stimulate phospholipase D. Natural ceramide 1-phosphate did not alter the cellular phosphorylation state of tyrosine residues or of mitogen-activated protein kinase. Furthermore, natural ceramide 1-phosphate failed to induce the expression of the proto-oncogenes c-myc and c-fos. Both the stimulation of DNA synthesis and the induction of proliferating-cell nuclear antigen by natural ceramide 1-phosphate were inhibited by natural ceramides. This work suggests that the use of methanol and dodecane to deliver natural ceramide 1-phosphate to cells may be useful for elucidation of the biological function(s) and mechanism(s) of action of ceramide 1-phosphate.

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Ubiquitinated proliferating cell nuclear antigen activates translesion DNA polymerases and REV1

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.0505949102 ◽

2005 ◽

Vol 102 (51) ◽

pp. 18361-18366 ◽

Cited By ~ 170

Author(s):

P. Garg ◽

P. M. Burgers

Keyword(s):

Proliferating Cell Nuclear Antigen ◽

Dna Polymerases ◽

Nuclear Antigen ◽

Translesion Dna ◽

Proliferating Cell

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A Single Domain in Human DNA Polymerase ι Mediates Interaction with PCNA: Implications for Translesion DNA Synthesis

Molecular and Cellular Biology ◽

10.1128/mcb.25.3.1183-1190.2005 ◽

2005 ◽

Vol 25 (3) ◽

pp. 1183-1190 ◽

Cited By ~ 39

Author(s):

Lajos Haracska ◽

Narottam Acharya ◽

Ildiko Unk ◽

Robert E. Johnson ◽

Jerard Hurwitz ◽

...

Keyword(s):

Dna Synthesis ◽

Tight Binding ◽

Dna Lesions ◽

Nuclear Antigen ◽

Binding Motifs ◽

Translesion Dna Synthesis ◽

Nucleotide Incorporation ◽

Y Family ◽

Stalled Replication Forks ◽

Proliferating Cell

ABSTRACT DNA polymerases (Pols) of the Y family rescue stalled replication forks by promoting replication through DNA lesions. Humans have four Y family Pols, η, ι, κ, and Rev1, of which Pols η, ι, and κ have been shown to physically interact with proliferating cell nuclear antigen (PCNA) and be functionally stimulated by it. However, in sharp contrast to the large increase in processivity that PCNA binding imparts to the replicative Pol, Polδ, the processivity of Y family Pols is not enhanced upon PCNA binding. Instead, PCNA binding improves the efficiency of nucleotide incorporation via a reduction in the apparent Km for the nucleotide. Here we show that Polι interacts with PCNA via only one of its conserved PCNA binding motifs, regardless of whether PCNA is bound to DNA or not. The mode of PCNA binding by Polι is quite unlike that in Polδ, where multisite interactions with PCNA provide for a very tight binding of the replicating Pol with PCNA. We discuss the implications of these observations for the accuracy of DNA synthesis during translesion synthesis and for the process of Pol exchange at the lesion site.

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