Further characterization of the yeastpso4-1 mutant: interaction withrad51 andrad52 mutants after photoinduced psoralen lesions

MoraisJunior, Marcos Antonio de; Vicente, Elisabete José; Brozmanová, J; Schenberg, Ana Clara Guerrini; Henriques, João Antônio Pêgas

doi:10.1007/bf02221550

Cited by 18 publications

(7 citation statements)

References 30 publications

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“…Of these ''PSO'' genes, seven have been demonstrated to have a role in DNA repair. Notably, in pso2-1͞snm1 and pso4-1 mutants, accumulation of DSBs was found by sucrose gradient analysis after interstrand crosslink-induced DNA damage, implicating their gene products in the processing and͞or rejoining of DNA ends (18,20,21). Genetic interactions have also been observed among yeast pso mutants, suggesting that their gene products may functionally interact to repair crosslink-induced DNA damage (18,20).…”

Section: Discussionmentioning

confidence: 95%

Role of human Pso4 in mammalian DNA repair and association with terminal deoxynucleotidyl transferase

Mahajan

Mitchell

2003

Proc. Natl. Acad. Sci. U.S.A.

100

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Terminal deoxynucleotidyl transferase (TdT; EC 2.7.7.31) adds nucleotides to DNA ends generated during V(D)J recombination that are subsequently processed by proteins involved in general doublestrand break (DSB) repair pathways. We report an association between TdT and a 55-kDa protein in lymphoid cells. This protein, identified as hPso4, is a homolog of the protein encoded by the PS04͞PRP19 gene in Saccharomyces cerevisiae that has pleiotropic functions in DNA recombination and error-prone repair. Purified hPso4 binds double-stranded DNA in a sequence-nonspecific manner but does not bind single-stranded DNA. hPso4 protein is induced 15-to 30-fold in cells by ␥ radiation and chemical mutagens but not by UV treatment. Loss of hPso4 expression induced by siRNA results in accumulation of DSBs, apoptosis, and decreased cell survival after DNA damage. We conclude that hPso4 plays a major and previously undefined role in mammalian DNA DSB repair.double-strand breaks ͉ end joining C ells encounter genomic lesions in the form of double-strand breaks (DSBs). In mammalian cells, DSB repair (DSBR) is mediated either by the error-free homologous recombination (HR) or the more predominant error-prone nonhomologous end joining (NHEJ) pathways (1). Agents that induce other types of DNA lesions, such as DNA interstrand crosslinks and adducts that proceed via DSB intermediates, are thought to invoke more than one type of DNA repair, including HR, nucleotide excision repair, and, to a lesser extent, the NHEJ pathway (2). The NHEJ repair proteins Ku70͞86, DNA PKcs, and Xrcc4͞ligase IV are ubiquitously expressed and function both in generalized DSBR and in the repair of ends generated during V(D)J recombination, a programmed gene rearrangement process that leads to the assembly of Ig and T cell receptors (3). Although these proteins seem sufficient for end joining in vitro, recent studies suggest the requirement of additional unknown factors for end joining in vivo (3, 4). One such recently identified human protein is artemis, mutations in which result in ionizing radiation sensitivity and severe combined immunodeficiency (5). Artemis protein has an inherent 5Ј to 3Ј singlestrand specific exonuclease activity. However, its association with and phosphorylation by DNA PKcs alter its activity to enable endonucleolytic processing of overhangs and cleavage of hairpin substrates such as those generated during V(D)J recombination (6).Terminal deoxynucleotidyl transferase (TdT; EC 2.7.7.31), a template-independent DNA polymerase, is unique in its expression and function, because it is expressed only in lymphoid cells undergoing V(D)J recombination (3) and has the ability to add nucleotides to the recombining V, (D), and J coding ends, thereby promoting antibody and receptor diversity (7). Our earlier studies revealed the association of TdT with the NHEJ protein Ku via the TdT BRCA1 C terminus (BRCT) region (8). Experiments performed with Ku86-deficient mice and cell lines have indeed demonstrated an absolute requirement of Ku86 for TdT-med...

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Section: Discussionmentioning

confidence: 95%

Role of human Pso4 in mammalian DNA repair and association with terminal deoxynucleotidyl transferase

Mahajan

Mitchell

2003

Proc. Natl. Acad. Sci. U.S.A.

100

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“…Previous reported findings including ours have indicated that Prp19 plays a direct role in DNA repair processing [1][2][3][4]20,21], although the nature of this role remains undefined. To examine whether hPrp19 self-association is affected by exposure of cells to DNA damaging agents, we expressed GFP-hPrp19, and treated the transfected cells with methyl-methane sulfonate (MMS).…”

Section: Dna Damage Affects Chromatin Association and The Structure Omentioning

confidence: 89%

“…Induced mutagenesis and induced and spontaneous mitotic recombination were all greatly reduced in this mutant suggesting that Pso4 is involved in a recombinational pathway of error-prone repair. Based on epistasis analysis PSO4 was assigned to both the yeast RAD6 and RAD52 groups indicating the pleiotropic nature of this mutation [3,4]. Interestingly, cloning of PSO4 showed that it was allelic to PRP19 a previously characterized component of the pre-mRNA splicing complex in both yeast and human cells [5][6][7][8][9][10].…”

Section: Introductionmentioning

confidence: 99%

The Prp19/Pso4 core complex undergoes ubiquitylation and structural alterations in response to DNA damage

Legerski²

2007

Biochemical and Biophysical Research Communications

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Prp19/Pso4, a U-box containing E3 ligase, has a demonstrated role in pre-mRNA splicing, but has also been implicated in both yeast and mammalian cells as having a direct role in DNA damage processing. In this report we provide further evidence in support of this latter assertion. We show that hPrp19 forms an ubiquitylated oligomeric species that is resistant to disruption by SDS gel electrophoresis under nonreducing conditions suggesting that is mediated by a thiolester between ubiquitin and a cysteine residue in Prp19. The level of this species is significantly enhanced upon treatment of cells with DNA damaging agents, and its association with chromatin is increased. In addition, hPrp19 is known to form a stable core complex with Cdc5L, Plrg1, and Spf27; however, ubiquitylated hPrp19 fails to interact with either Cdc5L or Plrg1 indicating that DNA damage can induce profound alterations to the hPrp19 core complex. Finally, we show that overexpression of hPrp19 in human cells provides a pro-survival affect in that it reduces the levels of apoptosis observed after exposure of cells to DNA damage.The pso4-1 mutant of S. cerevisiae was isolated in a genetic screen for strains sensitive to the cross-linking agent psoralen plus UVA (PUVA) [1,2]. Characterization of this mutant indicated that it was particularly sensitive to bifunctional alkylating agents, but was also sensitive to a broad range of DNA damaging agents including IR, UV, and monofuntional alkylating compounds. Induced mutagenesis and induced and spontaneous mitotic recombination were all greatly reduced in this mutant suggesting that Pso4 is involved in a recombinational pathway of error-prone repair. Based on epistasis analysis PSO4 was assigned to both the yeast RAD6 and RAD52 groups indicating the pleiotropic nature of this mutation [3,4]. Interestingly, cloning of PSO4 showed that it was allelic to PRP19 a previously characterized component of the pre-mRNA splicing complex in both yeast and human cells [5][6][7][8][9][10].The complete function of Prp19 in mRNA splicing is not known, however, it has been shown that the Prp19p-associated complex is required for activation of the pre-mRNA splicesome and for the stable association of the small nuclear RNAs U5 and U6 with the spliceosome after U4 is dissociated [11,12]. These findings suggest a possible structural role for the Prp19-associated complex in pre-mRNA splicing. Mammalian Prp19 associates with a large number of splicing factors, although, it appears to form a core complex with three other proteins including Cdc5L, Plrg1, and Spf27 [9]. The structure of the Prp19 core complex is not completely resolved, however, it has been shown that yeast Prp19 forms a tetramer via its coiled coil domains, and that the tetramer interacts with one copy of Cdc5L through the associated coiled coil domains *Corresponding author. Fax. 713-792-1474; E-Mail: rlegersk@mdanderson.org. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to ou...

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“…SNEV's involvement in DNA repair is consistent with the role of its yeast orthologue Pso4 (Prp19) ( 166 , 167 ), where the temperature-sensitive mutant strain pso4-1 displays a pleiotropic phenotype that includes sensitivity to 8-methoxy-psoralen/UVA treatment ( 168 ). In yeast, Pso4 has been assigned to epistasis groups rad6 and rad52, emphasizing its pleiotropic nature ( 169 , 170 ).…”

Section: Icl Repair Deficiency Contributes To Signs Of Accelerated Agmentioning

confidence: 99%

Contributions of DNA interstrand cross-links to aging of cells and organisms

Grillari‐Voglauer

Katinger

Voglauer

2007

Nucleic Acids Research

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Impaired DNA damage repair, especially deficient transcription-coupled nucleotide excision repair, leads to segmental progeroid syndromes in human patients as well as in rodent models. Furthermore, DNA double-strand break signalling has been pinpointed as a key inducer of cellular senescence. Several recent findings suggest that another DNA repair pathway, interstrand cross-link (ICL) repair, might also contribute to cell and organism aging. Therefore, we summarize and discuss here that (i) systemic administration of anti-cancer chemotherapeutics, in many cases DNA cross-linking drugs, induces premature progeroid frailty in long-term survivors; (ii) that ICL-inducing 8-methoxy-psoralen/UVA phototherapy leads to signs of premature skin aging as prominent long-term side effect and (iii) that mutated factors involved in ICL repair like ERCC1/XPF, the Fanconi anaemia proteins, WRN and SNEV lead to reduced replicative life span in vitro and segmental progeroid syndromes in vivo. However, since ICL-inducing drugs cause damage different from ICL and since all currently known ICL repair factors work in more than one pathway, further work will be needed to dissect the actual contribution of ICL damage to aging.

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Further characterization of the yeastpso4-1 mutant: interaction withrad51 andrad52 mutants after photoinduced psoralen lesions

Cited by 18 publications

References 30 publications

Role of human Pso4 in mammalian DNA repair and association with terminal deoxynucleotidyl transferase

Role of human Pso4 in mammalian DNA repair and association with terminal deoxynucleotidyl transferase

The Prp19/Pso4 core complex undergoes ubiquitylation and structural alterations in response to DNA damage

Contributions of DNA interstrand cross-links to aging of cells and organisms

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