DNA ligase I gene expression during differentiation and cell proliferation

Montecucco, Alessandra; Biamonti, Giuseppe; Savini, Elena; Focher, Federico; Spadari, Silvio; Ciarrocchi, Giovanni

doi:10.1093/nar/20.23.6209

Cited by 47 publications

(38 citation statements)

References 42 publications

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“…The extensive characterization of the replicative human DNA ligase I undertaken in the last decade demonstrated that the level of the protein strongly correlates with the rate of cell proliferation (Montecucco et al, 1992; 1995; Timson et al, 2000). Therefore we have investigated the possibility to use a new anti-DNA ligase I antibody (5H5) to detect proliferating cells.…”

Section: Discussionmentioning

confidence: 99%

“…Consistent with its role in DNA replication, DNA ligase I expression strongly correlates with the rate of cell proliferation. DNA ligase I activity raises up to 100 times in human lymphocytes upon PHA stimulation and both mRNA and protein level increase after serum stimulation of human and mouse primary fibroblasts (Montecucco et al, 1992;Pedrini et al, 1972). On the other hand a strong decrease of DNA ligase I gene expression is produced by confluence, serum starvation and cell differentiation (Camarda et al, 2004;Montecucco et al, 1992).…”

confidence: 99%

“…DNA ligase I activity raises up to 100 times in human lymphocytes upon PHA stimulation and both mRNA and protein level increase after serum stimulation of human and mouse primary fibroblasts (Montecucco et al, 1992;Pedrini et al, 1972). On the other hand a strong decrease of DNA ligase I gene expression is produced by confluence, serum starvation and cell differentiation (Camarda et al, 2004;Montecucco et al, 1992). In addition DNA ligase I expression is up-regulated during mouse liver-cell regeneration and in response to mitogenic stimuli (Gariboldi et al, 1995).…”

confidence: 99%

“…All reagents were from Sigma. Cells were grown at 37°C in a humidified atmosphere containing 5% CO2 and trypsinized when confluent.To obtain resting cells, confluent human primary fibroblasts were grown for 5 days in DMEM supplemented with 0.25% fetal calf serum as previously described (Montecucco et al, 1992). Peripheral blood lymphocytes were obtained from healthy donors.…”

Section: Cellsmentioning

confidence: 99%

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A new monoclonal antibody against DNA ligase I is a suitable marker of cell proliferation in cultured cell and tissue section samples

Vitolo

Mr²,

Montecucco

et al. 2009

Eur J Histochem

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The extensive characterization of the replicative human DNA ligase I (LigI) undertaken in the last decade demonstrated that the level of this protein strongly correlates with the rate of cell proliferation. This may allow to expand the repertoire of clinical biomarkers for the analysis of cell proliferation. We have produced a new monoclonal antibody (5H5) against LigI and exploited it as cell proliferation marker in Western blotting and immunofluorescence as well as in immunohistochemistry on paraffin tissue sections. The Western blot analysis showed that the LigI level detected by 5H5 antibody is high in all proliferating cells. On the contrary the protein is down regulated in resting human fibroblast and peripheral blood lymphocytes. Immunofluorescence analysis on cultured HeLa cells showed that 5H5 antibody labels all proliferating cells and displays the same staining pattern of BrdU in S-phase nuclei. Finally the analysis of serial sections of inflamed tonsils and NHL lymph nodes (either frozen or paraffin embedded) demonstrated that 5H5 marks the same population of cells as the Ki-67 antibody. Our results demonstrate that 5H5 antibody is a valuable tool for labeling proliferating cells that can be conveniently used in Western blotting, immunocytochemistry and immunohistochemistry.

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Section: Discussionmentioning

confidence: 99%

Section: Cellsmentioning

confidence: 99%

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A new monoclonal antibody against DNA ligase I is a suitable marker of cell proliferation in cultured cell and tissue section samples

Vitolo

Mr²,

Montecucco

et al. 2009

Eur J Histochem

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“…PCNA has also been identified as the central component of a targeting mechanism by which proteins that metabolize DNA locate their substrates (27). DNA ligase I is required for DNA-joining reactions that are an essential part of DNA replication and repair pathways (37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47)(48)(49). Recent characterization of the binding interaction between PCNA and DNA ligase I (30, 33) led us to investigate the effect of this binding on the ligation reaction.…”

Section: Discussionmentioning

confidence: 99%

DNA Ligase I and Proliferating Cell Nuclear Antigen Form a Functional Complex

Tom

Henricksen

Park

et al. 2001

Journal of Biological Chemistry

100

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DNA metabolism requires the coordinated activity of a multitude of enzymes and enzyme complexes. Although the initiation of DNA replication and DNA repair are regulated through different mechanisms, the reactions performed to complete these pathways are similar. In particular, Okazaki fragment processing (1) and long patch base excision repair (2, 3) share many enzymes needed for completion of these pathways. These include flap endonuclease 1 (FEN1), 1 proliferating cell nuclear antigen (PCNA), and DNA ligase I.During lagging strand DNA synthesis, numerous initiator RNA primers must be removed. The resulting gaps are filled in and sealed by ligation to complete DNA synthesis. Two nucleases, Dna2 and FEN1, are responsible for excising the RNA primer (4 -8). Both of these enzymes are unique structurespecific endonucleases. The preferred substrate contains a flap structure in which the RNA primer has been displaced to form a single-stranded tail (1, 9 -13). The flap structure probably arises as a result of displacement synthesis from an upstream Okazaki fragment by a complex of DNA polymerase ␦ and its accessory factors, PCNA and replication factor C (RFC) (14). Dna2 is thought to cleave beyond the RNA segment within the tail, and the remaining displaced DNA is removed by FEN1 (5-7). Finally, the two fragments are joined through ligation by DNA ligase I (1, 15).Long patch base excision repair utilizes several components common to Okazaki fragment processing to remove bases altered by ionizing radiation, oxidation, or alkylating agents (2, 3, 16 -21). During the repair process, an abasic site is generated after removal of a damaged base by a DNA N-glycosylase. An apurinic/apyrimidinic endonuclease subsequently cleaves on the 5Ј-side of the abasic sugar to create a nick within the DNA. Similar to the removal of initiator RNA primers, synthesis by a DNA polymerase lifts the damaged residue and a few additional downstream nucleotides to form a flap. As during replication, this structure is removed endonucleolytically by FEN1 followed by ligation of the resulting nick by DNA ligase I (2,3,17,19,21). This entire process is stimulated in the presence of PCNA (22).PCNA is a toroidal homotrimer that is assembled around double-stranded DNA to form a sliding clamp (23, 24). It has long been known to act as a processivity factor for DNA polymerases by tethering the polymerase to its template (25). However, PCNA also interacts with other replication proteins and appears to be responsible for recruiting these proteins to replication foci in vivo (26 -28). The interaction of PCNA and FEN1 has been examined extensively (12, 13, 29 -31). The FEN1 nuclease binds to the interdomain connecting loop region of PCNA (12,29,30,32), and this association leads to a potent stimulation of FEN1 cleavage activity (12,13,29). The physical interaction between the PCNA toroid and FEN1 enhances the binding stability of FEN1 to cleavage sites (13). In this way, PCNA serves to clamp FEN1 to its substrate in much the same way as this protein clamps DN...

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The N-terminal domain of human DNA ligase I contains the nuclear localization signal and directs the enzyme to sites of DNA replication.

et al. 1995

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DNA replication in mammalian cells occurs in discrete nuclear foci called ‘replication factories’. Here we show that DNA ligase I, the main DNA ligase activity in proliferating cells, associates with the factories during S phase but displays a diffuse nucleoplasmic distribution in non‐S phase nuclei. Immunolocalization analysis of both chloramphenicol acetyltransferase (CAT)‐DNA ligase I fusion proteins and epitope tagged DNA ligase I mutants allowed the identification of a 13 amino acid functional nuclear localization signal (NLS) located in the N‐terminal regulatory domain of the protein. Furthermore, the NLS is immediately preceded by a 115 amino acid region required for the association of the enzyme with the replication factories. We propose that in vivo the activity of DNA ligase I could be modulated through the control of its sub‐nuclear compartmentalization.

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DNA ligase I gene expression during differentiation and cell proliferation

Cited by 47 publications

References 42 publications

A new monoclonal antibody against DNA ligase I is a suitable marker of cell proliferation in cultured cell and tissue section samples

A new monoclonal antibody against DNA ligase I is a suitable marker of cell proliferation in cultured cell and tissue section samples

DNA Ligase I and Proliferating Cell Nuclear Antigen Form a Functional Complex

The N-terminal domain of human DNA ligase I contains the nuclear localization signal and directs the enzyme to sites of DNA replication.

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