UV-induced DNA incision and proliferating cell nuclear antigen recruitment to repair sites occur independently of p53–replication protein A interaction in p53 wild type and mutant ovarian carcinoma cells

DNA pol (polymerase) ϵ is thought to be the leading strand replicase in eukaryotes. In the present paper, we show that human DNA pol ϵ can efficiently bypass an 8-oxo-G (7,8-dihydro-8-oxoguanine) lesion on the template strand by inserting either dCMP or dAMP opposite to it, but it cannot bypass an abasic site. During replication, DNA pols associate with accessory proteins that may alter their bypass ability. We investigated the role of the human DNA sliding clamp PCNA (proliferating-cell nuclear antigen) and of the human single-stranded DNA-binding protein RPA (replication protein A) in the modulation of the DNA synthesis and translesion capacity of DNA pol ϵ. RPA inhibited the elongation by human DNA pol ϵ on templates annealed to short primers. PCNA did not influence the elongation by DNA pol ϵ and had no effect on inhibition of elongation caused by RPA. RPA inhibition was considerably reduced when the length of the primers was increased. On templates bearing the 8-oxo-G lesion, this inhibitory effect was more pronounced on DNA replication beyond the lesion, suggesting that RPA may prevent extension by DNA pol ϵ after incorporation opposite an 8-oxo-G. Neither PCNA nor RPA had any effect on the inability of DNA pol ϵ to replicate past the AP site, independent of the primer length.

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DNA Elongation by the Human DNA Polymerase λ Polymerase and Terminal Transferase Activities Are Differentially Coordinated by Proliferating Cell Nuclear Antigen and Replication Protein A

Journal of Biological Chemistry ◽

10.1074/jbc.m411650200 ◽

2004 ◽

Vol 280 (3) ◽

pp. 1971-1981 ◽

Cited By ~ 18

Author(s):

Giovanni Maga ◽

Kristijan Ramadan ◽

Giada. A. Locatelli ◽

Igor Shevelev ◽

Silvio Spadari ◽

...

Keyword(s):

Dna Polymerase ◽

Proliferating Cell Nuclear Antigen ◽

Protein A ◽

Replication Protein A ◽

Replication Protein ◽

Nuclear Antigen ◽

Terminal Transferase ◽

Human Dna ◽

Dna Polymerase Λ ◽

Proliferating Cell

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Cell Cycle Specific Plasmid DNA Replication in the Nuclear Extract ofSaccharomyces cerevisiae: Modulation by Replication Protein A and Proliferating Cell Nuclear Antigen†

Biochemistry ◽

10.1021/bi0120289 ◽

2002 ◽

Vol 41 (16) ◽

pp. 5255-5265 ◽

Cited By ~ 3

Author(s):

Atanaska V. Mitkova ◽

Esther E. Biswas ◽

Subhasis B. Biswas

Keyword(s):

Cell Cycle ◽

Dna Replication ◽

Plasmid Dna ◽

Proliferating Cell Nuclear Antigen ◽

Protein A ◽

Replication Protein A ◽

Nuclear Extract ◽

Replication Protein ◽

Nuclear Antigen ◽

Proliferating Cell

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Okazaki fragment processing: Modulation of the strand displacement activity of DNA polymerase by the concerted action of replication protein A, proliferating cell nuclear antigen, and flap endonuclease-1

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.251193198 ◽

2001 ◽

Vol 98 (25) ◽

pp. 14298-14303 ◽

Cited By ~ 73

Author(s):

G. Maga ◽

G. Villani ◽

V. Tillement ◽

M. Stucki ◽

G. A. Locatelli ◽

...

Keyword(s):

Dna Polymerase ◽

Proliferating Cell Nuclear Antigen ◽

Protein A ◽

Replication Protein A ◽

Nuclear Antigen ◽

Strand Displacement ◽

Concerted Action ◽

Flap Endonuclease 1 ◽

Okazaki Fragment Processing ◽

Proliferating Cell

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The N-degron protein degradation strategy for investigating the function of essential genes: requirement for replication protein A and proliferating cell nuclear antigen proteins for nucleotide excision repair in yeast extracts

Mutation Research/DNA Repair ◽

10.1016/s0921-8777(98)00031-7 ◽

1998 ◽

Vol 408 (3) ◽

pp. 183-194 ◽

Cited By ~ 10

Author(s):

Wenya Huang ◽

William J. Feaver ◽

Alan E. Tomkinson ◽

Errol C. Friedberg

Keyword(s):

Protein Degradation ◽

Nucleotide Excision Repair ◽

Proliferating Cell Nuclear Antigen ◽

Excision Repair ◽

Protein A ◽

Replication Protein A ◽

Essential Genes ◽

Nuclear Antigen ◽

Nucleotide Excision ◽

Proliferating Cell

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Characterization of the Proliferating Cell Nuclear Antigen of Leishmania donovani Clinical Isolates and Its Association with Antimony Resistance

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01847-13 ◽

2014 ◽

Vol 58 (6) ◽

pp. 2997-3007 ◽

Cited By ~ 9

Author(s):

Rati Tandon ◽

Sharat Chandra ◽

Rajendra Kumar Baharia ◽

Sanchita Das ◽

Pragya Misra ◽

...

Keyword(s):

Clinical Isolate ◽

Proliferating Cell Nuclear Antigen ◽

Leishmania Donovani ◽

Clinical Isolates ◽

Resistant Isolate ◽

Nuclear Antigen ◽

Wild Type ◽

Content Type ◽

Sensitive Isolate ◽

Proliferating Cell

ABSTRACTPreviously, through a proteomic analysis, proliferating cell nuclear antigen (PCNA) was found to be overexpressed in the sodium antimony gluconate (SAG)-resistant clinical isolate compared to that in the SAG-sensitive clinical isolate ofLeishmania donovani. The present study was designed to explore the potential role of the PCNA protein in SAG resistance inL. donovani. For this purpose, the protein was cloned, overexpressed, purified, and modeled. Western blot (WB) and real-time PCR (RT-PCR) analyses confirmed that PCNA was overexpressed by ≥3-fold in the log phase, stationary phase, and peanut agglutinin isolated procyclic and metacyclic stages of the promastigote form and by ∼5-fold in the amastigote form of the SAG-resistant isolate compared to that in the SAG-sensitive isolate.L. donovaniPCNA (LdPCNA) was overexpressed as a green fluorescent protein (GFP) fusion protein in a SAG-sensitive clinical isolate ofL. donovani, and modulation of the sensitivities of the transfectants to pentavalent antimonial (SbV) and trivalent antimonial (SbIII) drugs was assessedin vitroagainst promastigotes and intracellular (J774A.1 cell line) amastigotes, respectively. Overexpression of LdPCNA in the SAG-sensitive isolate resulted in an increase in the 50% inhibitory concentrations (IC50) of SbV(from 41.2 ± 0.6 μg/ml to 66.5 ± 3.9 μg/ml) and SbIII(from 24.0 ± 0.3 μg/ml to 43.4 ± 1.8 μg/ml). Moreover, PCNA-overexpressing promastigote transfectants exhibited less DNA fragmentation compared to that of wild-type SAG-sensitive parasites upon SbIIItreatment. In addition, SAG-induced nitric oxide (NO) production was found to be significantly inhibited in the macrophages infected with the transfectants compared with that in wild-type SAG-sensitive parasites. Consequently, we infer that LdPCNA has a significant role in SAG resistance inL. donovaniclinical isolates, which warrants detailed investigations regarding its mechanism.

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