scholarly journals p21CDKN1A Does Not Interfere with Loading of PCNA at DNA Replication Sites, but Inhibits Subsequent Binding of DNA Polymerase D at the G1/S Phase Transition

Cell Cycle ◽  
2003 ◽  
Vol 2 (6) ◽  
pp. 595-602 ◽  
Author(s):  
Ornella Cazzalini ◽  
Paola Perucca ◽  
Federica Riva ◽  
Lucia A. Stivala ◽  
Livia Bianchi ◽  
...  
Keyword(s):  
Phase Transition ◽  
Dna Replication ◽  
Dna Polymerase ◽  
S Phase ◽  
Replication Sites

scholarly journals A Coordinated Temporal Interplay of Nucleosome Reorganization Factor, Sister Chromatin Cohesion Factor, and DNA Polymerase α Facilitates DNA Replication

2004 ◽  
Vol 24 (21) ◽  
pp. 9568-9579 ◽  
Author(s):  
Yanjiao Zhou ◽  
Teresa S.-F. Wang
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
S Phase ◽  
Terminal Region ◽  
Replication Complex ◽  
Dna Polymerase Α ◽  
Chromatid Cohesion ◽  
Phase Progression

ABSTRACT DNA replication depends critically upon chromatin structure. Little is known about how the replication complex overcomes the nucleosome packages in chromatin during DNA replication. To address this question, we investigate factors that interact in vivo with the principal initiation DNA polymerase, DNA polymerase α (Polα). The catalytic subunit of budding yeast Polα (Pol1p) has been shown to associate in vitro with the Spt16p-Pob3p complex, a component of the nucleosome reorganization system required for both replication and transcription, and with a sister chromatid cohesion factor, Ctf4p. Here, we show that an N-terminal region of Polα (Pol1p) that is evolutionarily conserved among different species interacts with Spt16p-Pob3p and Ctf4p in vivo. A mutation in a glycine residue in this N-terminal region of POL1 compromises the ability of Pol1p to associate with Spt16p and alters the temporal ordered association of Ctf4p with Pol1p. The compromised association between the chromatin-reorganizing factor Spt16p and the initiating DNA polymerase Pol1p delays the Pol1p assembling onto and disassembling from the late-replicating origins and causes a slowdown of S-phase progression. Our results thus suggest that a coordinated temporal and spatial interplay between the conserved N-terminal region of the Polα protein and factors that are involved in reorganization of nucleosomes and promoting establishment of sister chromatin cohesion is required to facilitate S-phase progression.

scholarly journals Mrc1 and DNA Polymerase ɛ Function Together in Linking DNA Replication and the S Phase Checkpoint

2008 ◽  
Vol 32 (1) ◽  
pp. 106-117 ◽  
Author(s):  
Huiqiang Lou ◽  
Makiko Komata ◽  
Yuki Katou ◽  
Zhiyun Guan ◽  
Clara C. Reis ◽  
...  
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
S Phase ◽  
S Phase Checkpoint

scholarly journals Cell cycle-dependent phosphorylation and dephosphorylation of the yeast DNA polymerase alpha-primase B subunit.

1995 ◽  
Vol 15 (2) ◽  
pp. 883-891 ◽  
Author(s):  
M Foiani ◽  
G Liberi ◽  
G Lucchini ◽  
P Plevani
Keyword(s):  
Cell Cycle ◽  
Dna Replication ◽  
Dna Polymerase ◽  
Posttranslational Modifications ◽  
S Phase ◽  
Dna Polymerase Alpha ◽  
B Subunit ◽  
Initiation Of Dna Replication ◽  
Cycle Dependent

The yeast DNA polymerase alpha-primase B subunit functions in initiation of DNA replication. This protein is present in two forms, of 86 and 91 kDa, and the p91 polypeptide results from cell cycle-regulated phosphorylation of p86. The B subunit present in G1 arises by dephosphorylation of p91 while cells are exiting from mitosis, becomes phosphorylated in early S phase, and is competent and sufficient to initiate DNA replication. The B subunit transiently synthesized as a consequence of periodic transcription of the POL12 gene is phosphorylated no earlier than G2. Phosphorylation of the B subunit does not require execution of the CDC7-dependent step and ongoing DNA synthesis. We suggest that posttranslational modifications of the B subunit might modulate the role of DNA polymerase alpha-primase in DNA replication.

scholarly journals Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites.

1987 ◽  
Vol 105 (4) ◽  
pp. 1549-1554 ◽  
Author(s):  
R Bravo ◽  
H Macdonald-Bravo
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
S Phase ◽  
Nuclear Antigen ◽  
Dna Polymerase Delta ◽  
Quiescent Cells ◽  
Replication Sites ◽  
Nuclear Structures ◽  
Proliferating Cell ◽  
Two Populations

Pulse-chase experiments have revealed that cyclin, the auxiliary protein of DNA polymerase-delta, is stable during the transition from growth to quiescence in 3T3 cells. Immunoblotting together with immunofluorescence analysis has shown that the amount of cyclin after 24 h of quiescence is 30-40% of that of growing cells and that it presents a nucleoplasmic staining. Immunofluorescence studies show the existence of two populations of cyclin during the S phase, one that is nucleoplasmic as in quiescent cells and is easily extracted by detergent, and another that is associated to specific nuclear structures. By using antibromodeoxyuridine immunofluorescence to detect the sites of DNA synthesis, it was shown that the staining patterns of the replicon clusters and their order of appearance throughout the S phase are identical to those observed for cyclin. Two-dimensional gel analysis of Triton-extracted cells show that 20-30% of cyclin remains associated with the replicon clusters. This population of cyclin could not be released from the nucleus using high-salt extractions. This demonstrates that cyclin is tightly associated to the sites of DNA replication and that it must have a fundamental role in DNA synthesis in eukaryotic cells.

scholarly journals Initiation and termination of DNA replication during S phase in relation to cyclins D1, E and A, p21WAF1, Cdt1 and the p12 subunit of DNA polymerase δ revealed in individual cells by cytometry

Oncotarget ◽  
2015 ◽  
Vol 6 (14) ◽  
pp. 11735-11750 ◽  
Author(s):  
Zbigniew Darzynkiewicz ◽  
Hong Zhao ◽  
Sufang Zhang ◽  
Marietta Y.W.T. Lee ◽  
Ernest Y.C. Lee ◽  
...  
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
S Phase ◽  
Dna Polymerase Δ

scholarly journals Contribution of DNA polymerase δ to DNA replication in permeable CHO cells synchronized in S phase

1989 ◽  
Vol 17 (12) ◽  
pp. 4757-4767 ◽  
Author(s):  
Alexei Basnakian ◽  
Gaspar Banfalvi ◽  
Nilima Sarkar
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Cho Cells ◽  
S Phase ◽  
Dna Polymerase Δ

scholarly journals Discrimination of Cell Nuclei in Early S-phase, Mid-to-late S-phase, and G2/M-phase by Sequential Administration of 5-Bromo-2'-Deoxyuridine and 5-Chloro-2'-Deoxyuridine

2005 ◽  
Vol 53 (11) ◽  
pp. 1365-1370 ◽  
Author(s):  
Kumiko Yamada ◽  
Reiji Semba ◽  
XiaoHui Ding ◽  
Ning Ma ◽  
Masato Nagahama
Keyword(s):  
Monoclonal Antibody ◽  
Dna Replication ◽  
S Phase ◽  
Phase Identification ◽  
Cell Nuclei ◽  
Intracellular Movement ◽  
Sequential Administration ◽  
M Phase ◽  
Replication Sites ◽  
Small Intestines

5-Bromo-2'-deoxyuridine (BrdU) and 5-chloro-2'-deoxyuridine (CldU) were sequentially administered intraperitoneally into mice at 1-hr intervals. After one additional hr, the small intestines were resected, fixed, and embedded in paraffin. In histological sections stained with monoclonal antibody Br-3 reactive to both BrdU and CldU, and CldU antibody reactive only to CldU, three types of staining could be identified in the proliferating zone. Cells with nuclei stained only with Br-3 antibody were estimated to have completed DNA replication during the first 1 hr and were fixed in G2/M-phase. Those nuclei were frequently found in apical areas of the simple columnar epithelium of the intestine, whereas other nuclei were located basally in the cells. This observation suggested intracellular movement of cell nuclei in G2/M-phase. Identification of cells in early S-phase became possible using these antibodies in combination with DAB and fluorescence stainings. Replication sites in early S-phase nuclei were found to be numerous, whereas in late S-phase they were larger in size and much smaller in number.

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