scholarly journals Caspase-2 activation in the absence of PIDDosome formation

2009 ◽  
Vol 185 (2) ◽  
pp. 291-303 ◽  
Author(s):  
Claudia Manzl ◽  
Gerhard Krumschnabel ◽  
Florian Bock ◽  
Benedicte Sohm ◽  
Verena Labi ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Dna Damage ◽  
High Molecular Weight ◽  
Nuclear Translocation ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Death Domain ◽  
High Molecular Weight Complex ◽  
Effector Caspase ◽  
Caspase 2

PIDD (p53-induced protein with a death domain [DD]), together with the bipartite adapter protein RAIDD (receptor-interacting protein-associated ICH-1/CED-3 homologous protein with a DD), is implicated in the activation of pro–caspase-2 in a high molecular weight complex called the PIDDosome during apoptosis induction after DNA damage. To investigate the role of PIDD in cell death initiation, we generated PIDD-deficient mice. Processing of caspase-2 is readily detected in the absence of PIDDosome formation in primary lymphocytes. Although caspase-2 processing is delayed in simian virus 40–immortalized pidd−/− mouse embryonic fibroblasts, it still depends on loss of mitochondrial integrity and effector caspase activation. Consistently, apoptosis occurs normally in all cell types analyzed, suggesting alternative biological roles for caspase-2 after DNA damage. Because loss of either PIDD or its adapter molecule RAIDD did not affect subcellular localization, nuclear translocation, or caspase-2 activation in high molecular weight complexes, we suggest that at least one alternative PIDDosome-independent mechanism of caspase-2 activation exists in mammals in response to DNA damage.

scholarly journals Integration of simian virus 40 into cellular DNA occurs at or near topoisomerase II cleavage hot spots induced by VM-26 (teniposide).

1993 ◽  
Vol 13 (10) ◽  
pp. 6190-6200 ◽  
Author(s):  
A L Bodley ◽  
H C Huang ◽  
C Yu ◽  
L F Liu
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Hot Spots ◽  
Topoisomerase Ii ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
High Molecular Weight Form ◽  
Cellular Dna ◽  
Sv40 Dna ◽  
Molecular Weight Form

Inhibition of DNA topoisomerase II in simian virus 40 (SV40)-infected BSC-1 cells with a topoisomerase II poison, VM-26 (teniposide), resulted in rapid conversion of a population of the SV40 DNA into a high-molecular-weight form. Characterization of this high-molecular-weight form of SV40 DNA suggests that it is linear, double stranded, and a recombinant with SV40 DNA sequences covalently joined to cellular DNA. The majority of the integrants contain fewer than two tandem copies of SV40 DNA. Neither DNA-damaging agents, such as mitomycin and UV, nor the topoisomerase I inhibitor camptothecin induced detectable integration in this system. In addition, the recombination junctions within the SV40 portion of the integrants correlate with VM-26-induced, topoisomerase II cleavage hot spots on SV40 DNA. These results suggest a direct and specific role for topoisomerase II and possibly the enzyme-inhibitor-DNA ternary cleavable complex in integration. The propensity of poisoned topoisomerase II to induce viral integration also suggests a role for topoisomerase II in a pathway of chromosomal DNA rearrangements.

scholarly journals DNA damage stabilizes interaction of CSB with the transcription elongation machinery

2004 ◽  
Vol 166 (1) ◽  
pp. 27-36 ◽  
Author(s):  
Vincent van den Boom ◽  
Elisabetta Citterio ◽  
Deborah Hoogstraten ◽  
Angelika Zotter ◽  
Jean-Marc Egly ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Dna Damage ◽  
Dna Repair ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
High Molecular Weight ◽  
Transcription Elongation ◽  
Cockayne Syndrome ◽  
Dna Lesions ◽  
High Molecular Weight Complex

The Cockayne syndrome B (CSB) protein is essential for transcription-coupled DNA repair (TCR), which is dependent on RNA polymerase II elongation. TCR is required to quickly remove the cytotoxic transcription-blocking DNA lesions. Functional GFP-tagged CSB, expressed at physiological levels, was homogeneously dispersed throughout the nucleoplasm in addition to bright nuclear foci and nucleolar accumulation. Photobleaching studies showed that GFP-CSB, as part of a high molecular weight complex, transiently interacts with the transcription machinery. Upon (DNA damage-induced) transcription arrest CSB binding these interactions are prolonged, most likely reflecting actual engagement of CSB in TCR. These findings are consistent with a model in which CSB monitors progression of transcription by regularly probing elongation complexes and becomes more tightly associated to these complexes when TCR is active.

Integration of simian virus 40 into cellular DNA occurs at or near topoisomerase II cleavage hot spots induced by VM-26 (teniposide)

1993 ◽  
Vol 13 (10) ◽  
pp. 6190-6200
Author(s):  
A L Bodley ◽  
H C Huang ◽  
C Yu ◽  
L F Liu
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Hot Spots ◽  
Topoisomerase Ii ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
High Molecular Weight Form ◽  
Cellular Dna ◽  
Sv40 Dna ◽  
Molecular Weight Form

Inhibition of DNA topoisomerase II in simian virus 40 (SV40)-infected BSC-1 cells with a topoisomerase II poison, VM-26 (teniposide), resulted in rapid conversion of a population of the SV40 DNA into a high-molecular-weight form. Characterization of this high-molecular-weight form of SV40 DNA suggests that it is linear, double stranded, and a recombinant with SV40 DNA sequences covalently joined to cellular DNA. The majority of the integrants contain fewer than two tandem copies of SV40 DNA. Neither DNA-damaging agents, such as mitomycin and UV, nor the topoisomerase I inhibitor camptothecin induced detectable integration in this system. In addition, the recombination junctions within the SV40 portion of the integrants correlate with VM-26-induced, topoisomerase II cleavage hot spots on SV40 DNA. These results suggest a direct and specific role for topoisomerase II and possibly the enzyme-inhibitor-DNA ternary cleavable complex in integration. The propensity of poisoned topoisomerase II to induce viral integration also suggests a role for topoisomerase II in a pathway of chromosomal DNA rearrangements.

P4-129 Genetic study of APH-1 and PEN-2, components of presenilin high molecular weight complex

2004 ◽  
Vol 25 ◽  
pp. S512
Author(s):  
Toshitaka Kawarai ◽  
Antonio Orlacchio ◽  
Ekaterina Rogaeva ◽  
Susan Ling ◽  
Hiroshi Hasegawa ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Genetic Study ◽  
High Molecular Weight Complex

scholarly journals Binding of a sequence-specific single-stranded DNA-binding factor to the simian virus 40 core origin inverted repeat domain is cell cycle regulated.

1993 ◽  
Vol 13 (1) ◽  
pp. 408-420 ◽  
Author(s):  
E P Carmichael ◽  
J M Roome ◽  
A F Wahl
Keyword(s):  
Cell Cycle ◽  
Dna Damage ◽  
Dna Binding ◽  
Dna Synthesis ◽  
Inverted Repeat ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Single Stranded Dna ◽  
Repeat Domain

The inverted repeat domain (IR domain) within the simian virus 40 origin of replication is the site of initial DNA melting prior to the onset of DNA synthesis. The domain had previously been shown to be bound by a cellular factor in response to DNA damage. We demonstrate that two distinct cellular components bind opposite strands of the IR domain. Replication protein A (RPA), previously identified as a single-stranded DNA binding protein required for origin-specific DNA replication in vitro, is shown to have a preference for the pyrimidine-rich strand. A newly described component, IR factor B (IRF-B), specifically recognizes the opposite strand. IRF-B binding activity in nuclear extract varies significantly with cell proliferation and the cell cycle, so that binding of IRF-B to the IR domain is negatively correlated with the onset of DNA synthesis. Loss of IRF-B binding from the nucleus also occurs in response to cellular DNA damage. UV cross-linking indicates that the core binding component of IRF-B is a protein of ca. 34 kDa. We propose that RPA and IRF-B bind opposite strands of the IR domain and together may function in the regulation of origin activation.

scholarly journals High molecular weight complex of tyrosyl-tRNA synthetase from bovine liver

1991 ◽  
Vol 7 (1) ◽  
pp. 63-69 ◽  
Author(s):  
D. V. Gnatenko ◽  
A. I. Kornelyuk ◽  
I. V. Kurochkin ◽  
G. H. Matsuka
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Bovine Liver ◽  
Trna Synthetase ◽  
High Molecular Weight Complex

scholarly journals Multiple DNA Damage Signaling and Repair Pathways Deregulated by Simian Virus 40 Large T Antigen

2010 ◽  
Vol 84 (16) ◽  
pp. 8007-8020 ◽  
Author(s):  
Sergei Boichuk ◽  
Liang Hu ◽  
Jennifer Hein ◽  
Ole V. Gjoerup
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Viral Replication ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Productive Infection ◽  
Large T Antigen ◽  
Sv40 Large T Antigen ◽  
Viral Origin

ABSTRACT We demonstrated previously that expression of simian virus 40 (SV40) large T antigen (LT), without a viral origin, is sufficient to induce the hallmarks of a cellular DNA damage response (DDR), such as focal accumulation of γ-H2AX and 53BP1, via Bub1 binding. Here we expand our characterization of LT effects on the DDR. Using comet assays, we demonstrate that LT induces overt DNA damage. The Fanconi anemia pathway, associated with replication stress, becomes activated, since FancD2 accumulates in foci, and monoubiquitinated FancD2 is detected on chromatin. LT also induces a distinct set of foci of the homologous recombination repair protein Rad51 that are colocalized with Nbs1 and PML. The FancD2 and Rad51 foci require neither Bub1 nor retinoblastoma protein binding. Strikingly, wild-type LT is localized on chromatin at, or near, the Rad51/PML foci, but the LT mutant in Bub1 binding is not localized there. SV40 infection was previously shown to trigger ATM activation, which facilitates viral replication. We demonstrate that productive infection also triggers ATR-dependent Chk1 activation and that Rad51 and FancD2 colocalize with LT in viral replication centers. Using small interfering RNA (siRNA)-mediated knockdown, we demonstrate that Rad51 and, to a lesser extent, FancD2 are required for efficient viral replication in vivo, suggesting that homologous recombination is important for high-level extrachromosomal replication. Taken together, the interplay of LT with the DDR is more complex than anticipated, with individual domains of LT being connected to different subcomponents of the DDR and repair machinery.

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