Role of the second-largest subunit of DNA polymerase α in the interaction between the catalytic subunit and hyperphosphorylated retinoblastoma protein in late S phase

2006 ◽  
Vol 1764 (9) ◽  
pp. 1447-1453 ◽  
Author(s):  
Masaharu Takemura ◽  
Shonen Yoshida ◽  
Tetsu Akiyama ◽  
Masatoshi Kitagawa ◽  
Yoshiji Yamada
Keyword(s):  
Dna Polymerase ◽  
Retinoblastoma Protein ◽  
S Phase ◽  
Catalytic Subunit ◽  
Dna Polymerase Α

scholarly journals A Mutation in DNA Polymerase α Rescues WEE1KO Sensitivity to HU

2021 ◽  
Vol 22 (17) ◽  
pp. 9409
Author(s):  
Thomas Eekhout ◽  
José Antonio Pedroza-Garcia ◽  
Pooneh Kalhorzadeh ◽  
Geert De Jaeger ◽  
Lieven De Veylder
Keyword(s):  
Cell Cycle ◽  
Dna Replication ◽  
Dna Polymerase ◽  
Driving Force ◽  
S Phase ◽  
Catalytic Subunit ◽  
Genomic Integrity ◽  
Cyclin Dependent Kinases ◽  
Dna Polymerase Α ◽  
Wee1 Kinase

During DNA replication, the WEE1 kinase is responsible for safeguarding genomic integrity by phosphorylating and thus inhibiting cyclin-dependent kinases (CDKs), which are the driving force of the cell cycle. Consequentially, wee1 mutant plants fail to respond properly to problems arising during DNA replication and are hypersensitive to replication stress. Here, we report the identification of the polα-2 mutant, mutated in the catalytic subunit of DNA polymerase α, as a suppressor mutant of wee1. The mutated protein appears to be less stable, causing a loss of interaction with its subunits and resulting in a prolonged S-phase.

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.

Role of DNA polymerase α and δ in radiation clastogenesis

1991 ◽  
Vol 262 (1) ◽  
pp. 31-36 ◽  
Author(s):  
Michael A. Bender ◽  
Ruth C. Moore ◽  
Beatrice E. Pyatt
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerase Α

scholarly journals The Saccharomyces telomere-binding protein Cdc13p interacts with both the catalytic subunit of DNA polymerase α and the telomerase-associated Est1 protein

2000 ◽  
Vol 14 (14) ◽  
pp. 1777-1788 ◽  
Author(s):  
Haiyan Qi ◽  
Virginia A. Zakian
Keyword(s):  
Dna Polymerase ◽  
Binding Protein ◽  
Point Mutations ◽  
Single Strand ◽  
Catalytic Subunit ◽  
Full Length ◽  
Temperature Sensitive ◽  
Dna Polymerase Α ◽  
Telomere Binding Protein

Saccharomyces telomeres consist of ∼350 bp of C1-3A/TG1-3 DNA. Most of this ∼350 bp is replicated by standard, semiconservative DNA replication. After conventional replication, the C1-3A strand is degraded to generate a long single strand TG1-3 tail that can serve as a substrate for telomerase. Cdc13p is a single strand TG1-3DNA-binding protein that localizes to telomeres in vivo. Genetic data suggest that the Cdc13p has multiple roles in telomere replication. We used two hybrid analysis to demonstrate that Cdc13p interacted with both the catalytic subunit of DNA polymerase α, Pol1p, and the telomerase RNA-associated protein, Est1p. The association of these proteins was confirmed by biochemical analysis using full-length or nearly full-length proteins. Point mutations in either CDC13 orPOL1 that reduced the Cdc13p–Pol1p interaction resulted in telomerase mediated telomere lengthening. Over–expression of the carboxyl terminus of Est1p partially suppressed the temperature sensitive lethality of a cdc13-1 strain. We propose that Cdc13p's interaction with Est1p promotes TG1-3 strand lengthening by telomerase and its interaction with Pol1p promotes C1-3A strand resynthesis by DNA polymerase α.

scholarly journals Molecular cloning of the cDNA for the catalytic subunit of plant DNA polymerase α and its cell-cycle dependent expression

1997 ◽  
Vol 2 (11) ◽  
pp. 695-709 ◽  
Author(s):  
Masayuki Yokoi ◽  
Masaki Ito ◽  
Masako Izumi ◽  
Hiroshi Miyazawa ◽  
Hirokazu Nakai ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Molecular Cloning ◽  
Dna Polymerase ◽  
Catalytic Subunit ◽  
Dna Polymerase Α ◽  
Plant Dna ◽  
Cycle Dependent

Regulation of the synthesis of DNA polymerase-α in regenerating liver by calcium and 1,25-dihydroxyvitamin D3

1989 ◽  
Vol 67 (7) ◽  
pp. 345-351 ◽  
Author(s):  
Marianna Sikorska ◽  
Ian de Belle ◽  
James F. Whitfield ◽  
P. Roy Walker
Keyword(s):  
Dna Polymerase ◽  
Calcium Ions ◽  
Nuclear Matrix ◽  
Critical Stage ◽  
Dihydroxyvitamin D3 ◽  
Dna Polymerase Α ◽  
Enzyme Molecules ◽  
Rat Livers ◽  
Α Activity

Digestion of nuclei from normal or partially hepatectomised rat livers with endogenous nucleases liberated a pool of cryptic DNA polymerase-α activity which had previously gone unrecognised. Most of this activity is released into the supernatant as free enzyme molecules (11S), but a small fraction of it is released as a complex of 16S (probably with DNA primase). About 40% of the enzyme remains in the pellet, which contains undigested chromatin and components of the residual nuclear matrix and nucleoskeletal structures. Virtually all of this remaining activity is extracted by 2.0 M salt. The activity of DNA polymerase-α increases equally in all nuclear fractions during the period of DNA replication in regenerating rat liver. Lowering of the serum calcium level by thyroparathyroidectomy does not affect basal DNA polymerase-α activity, but prevents induction of the enzyme during the later stages of prereplicative development. However, an injection of 1α,25-dihydroxy vitamin D3 into the rat during the first 6 h after partial hepatectomy restores the ability of the hepatocytes to induce DNA polymerase-α activity and initiate DNA synthesis. These results are discussed in terms of the role of calcium ions in the regulation of the critical stage of prereplicative development which commits the cells to DNA replication.Key words: replication, nuclear, matrix, complex, lamina, reductase.

scholarly journals Essential Role of Sna41/Cdc45 in Loading of DNA Polymerase α onto Minichromosome Maintenance Proteins in Fission Yeast

2001 ◽  
Vol 276 (28) ◽  
pp. 26189-26196 ◽  
Author(s):  
Masashi Uchiyama ◽  
Dominic Griffiths ◽  
Ken-ichi Arai ◽  
Hisao Masai
Keyword(s):  
Fission Yeast ◽  
Dna Polymerase ◽  
Essential Role ◽  
Minichromosome Maintenance ◽  
Dna Polymerase Α

scholarly journals Interaction of the retinoblastoma protein (pRb) with the catalytic subunit of DNA polymerase δ (p125)

Oncogene ◽  
2000 ◽  
Vol 19 (48) ◽  
pp. 5464-5470 ◽  
Author(s):  
Nancy A Krucher ◽  
Adam Zygmunt ◽  
Nayef Mazloum ◽  
Sama Tamrakar ◽  
John W Ludlow ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Retinoblastoma Protein ◽  
Catalytic Subunit ◽  
Dna Polymerase Δ

scholarly journals The Second-Largest Subunit of the Mouse DNA Polymerase α-Primase Complex Facilitates Both Production and Nuclear Translocation of the Catalytic Subunit of DNA Polymerase α

1998 ◽  
Vol 18 (6) ◽  
pp. 3552-3562 ◽  
Author(s):  
Takeshi Mizuno ◽  
Nobutoshi Ito ◽  
Masayuki Yokoi ◽  
Akio Kobayashi ◽  
Katsuyuki Tamai ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Mammalian Cells ◽  
Nuclear Translocation ◽  
Expression System ◽  
Physiological Role ◽  
Polymerase Activity ◽  
Catalytic Subunit ◽  
Dna Polymerase Α ◽  
Localization Signal ◽  
Primase Complex

ABSTRACT DNA polymerase α-primase is a replication enzyme necessary for DNA replication in all eukaryotes examined so far. Mouse DNA polymerase α is made up of four subunits, the largest of which is the catalytic subunit with a molecular mass of 180 kDa (p180). This subunit exists as a tight complex with the second-largest subunit (p68), whose physiological role has remained unclear up until now. We set out to characterize these subunits individually or in combination by using a cDNA expression system in cultured mammalian cells. Coexpression of p68 markedly increased the protein level of p180, with the result that ectopically generated DNA polymerase activity was dramatically increased. Immunofluorescence analysis showed that while either singly expressed p180 or p68 was localized in the cytoplasm, cotransfection of both subunits resulted in colocalization in the nucleus. We identified a putative nuclear localization signal for p180 (residues 1419 to 1437) and found that interaction with p68 is essential for p180 to translocate into the nucleus. These results indicate that association of p180 with p68 is important for both protein synthesis of p180 and translocation into the nucleus, implying that p68 plays a pivotal role in the newly synthesized DNA polymerase α complex.

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