Selective Cleavage of Human DNA: RecA-Assisted Restriction Endonuclease (RARE) Cleavage

Ferrin, Lance J.; Camerini‐Otero, R. Daniel

doi:10.1126/science.1962209

Cited by 201 publications

(110 citation statements)

References 19 publications

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“…Following incubation of the DMC1 presynaptic filament with RAD51AP1, linear duplex DNA that harbors a homologous target, including an embedded SspI restriction site, is added. Formation of the synaptic complex, wherein the presynaptic filament is homologously paired to the target duplex region, results in protection of the duplex DNA against digestion by the restriction enzyme SspI (25,29,37). As reported before (29), neither the DMC1 presynaptic filament nor RAD51AP1 afforded significant protection against SspI (Fig.…”

Section: Resultsmentioning

confidence: 61%

RAD51-associated Protein 1 (RAD51AP1) Interacts with the Meiotic Recombinase DMC1 through a Conserved Motif

Dunlop

Dray

Zhao

et al. 2011

Journal of Biological Chemistry

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Background: RAD51AP1 physically and functionally interacts with the RAD51 and DMC1 recombinases. Results: Mutational analysis showed that the WVPP sequence in RAD51AP1 is part of the DMC1-specific interaction motif. Conclusion: RAD51AP1 interacts with RAD51 and DMC1 through distinct motifs. Significance: RAD51AP1 likely functions in meiotic homologous recombination by enhancing the recombinase activity of both RAD51 and DMC1.

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Section: Resultsmentioning

confidence: 61%

RAD51-associated Protein 1 (RAD51AP1) Interacts with the Meiotic Recombinase DMC1 through a Conserved Motif

Dunlop

Dray

Zhao

et al. 2011

Journal of Biological Chemistry

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“…We submit that when DNA damages become extensive, the intracellular assemblies progressively assume a protective role, acting to physically shield the DNA molecules. Double-stranded DNA segments within RecA-mediated homologous joints were indeed shown to be protected against DNAmodifying enzymes (35). Moreover, complete degradation of chromosomes was observed in ⌬RecA cells (6).…”

Section: Discussionmentioning

confidence: 86%

Ordered intracellular RecA–DNA assemblies: A potential site of in vivo RecA-mediated activities

Levin-Zaidman

Frenkiel-Krispin

Shimoni

et al. 2000

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

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The inducible SOS response increases the ability of bacteria to cope with DNA damage through various DNA repair processes in which the RecA protein plays a central role. Here we present the first study of the morphological aspects that accompany the SOS response in Escherichia coli. We find that induction of the SOS system in wild-type bacteria results in a fast and massive intracellular coaggregation of RecA and DNA into a lateral macroscopic assembly. The coaggregates comprise substantial portions of both the cellular RecA and the DNA complement. The structural features of the coaggregates and their relation to in vitro RecA-DNA networks, as well as morphological studies of strains carrying RecA mutants, are all consistent with the possibility that the intracellular assemblies represent a functional entity in which RecA-mediated DNA repair and protection activities occur.biocrystallization ͉ DNA repair ͉ homology search ͉ stress response R ecA-mediated DNA recombination and repair processes proceed through several sequential phases (1-3). A presynaptic filament in which RecA molecules coat a single-stranded DNA substrate is initially formed. The filament acts then as a sequence-specific DNA-binding entity, capable of searching and binding dsDNA sites that are homologous to the RecA-coated segment. Within the resulting joint species, DNA strand exchange and heteroduplex extension processes are promoted. The mechanism that enables a rapid search for DNA homology in vivo, within a highly crowded and complex genome, remains enigmatic.To reach its target, any sequence-specific DNA-binding protein must overcome two general obstacles: a minute cellular concentration of the target, and a vast excess of nontarget, yet still competitive, DNA sites. The search for a homologous DNA site conducted by the RecA-DNA presynaptic filament shares these hurdles, but is further encumbered by the uniquely adverse diffusion characteristics of its components. A DNA target corresponds to a segment that is part of, and embedded within, the chromosome. This, and the large structural asymmetry of DNA, conspire to minimize the diffusion constant of DNA sites (4, 5). In a homology search executed by the RecA-DNA filament, both the searching and the target entities are chromosomal DNA sites whose small diffusion constants drastically attenuate their encounter rate. How then does a RecA-mediated intracellular search evade the kinetic impediments that are intrinsic to the nature of its components?Here we show that damages inflicted on bacterial DNA lead to a rapid formation of an ordered intracellular assembly that accommodates both RecA and DNA. We suggest that the striated morphology of this RecA-DNA assembly is capable of promoting an in vivo homology search by attenuating both the sampling volume and the dimensionality of the process. Moreover, RecA was shown to protect chromosomal DNA from degradation (6) through unknown mechanisms. The tight crystalline packaging that is progressively assumed by the intracellular RecA-DNA assemblies as ...

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“…1). Cosmids A6 and SN5, containing a copy of the complete coding sequence of the MDV unique gene meq, were used for the deletion of this gene by the RecA-assisted restriction endonuclease cleavage method (25). Briefly, two oligonucleotides, Meq Taq3Ј (5Ј-TTT ATG TCA GTA AAT CGA TAA ATA ATG CCT TT-3Јpositions 5,589-5,620 and 136,000-136,031) and Meq Taq5Ј (5Ј-ACG ATC CGT CCC CCC TCG ATC TTT CTC TCG GGT CG-3Ј positions 6,673-6,707 and 134,913-134,947), located at both ends of the meq gene, were used to protect the TaqI sites (positions 5,603, 6,672, and 6,689 in SN5 cosmid and positions 134,928, 134,946, and 136,014 in the A6 cosmid) from methylation.…”

mentioning

confidence: 99%

Marek's disease virus-encoded Meq gene is involved in transformation of lymphocytes but is dispensable for replication

Lupiani

Lee

Chen

et al. 2004

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

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149

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Marek's disease virus (MDV) causes an acute lymphoproliferative disease in chickens, resulting in T cell lymphomas in visceral organs and peripheral nerves. Earlier studies have determined that the repeat regions of oncogenic serotype 1 MDV encode a basic leucine zipper protein, Meq, which structurally resembles the Jun͞Fos family of transcriptional activators. Meq is consistently expressed in MDV-induced tumor cells and has been suggested as the MDVassociated oncogene. To study the function of Meq, we have generated an rMd5⌬Meq virus by deleting both copies of the meq gene from the genome of a very virulent strain of MDV. Growth curves in cultured fibroblasts indicated that Meq is dispensable for in vitro virus replication. In vivo replication in lymphoid organs and feather follicular epithelium was also not impaired, suggesting that Meq is dispensable for lytic infection in chickens. Reactivation of the rMd5⌬Meq virus from peripheral blood lymphocytes was reduced, suggesting that Meq is involved but not essential for latency. Pathogenesis experiments showed that the rMd5⌬Meq virus was fully attenuated in chickens because none of the infected chickens developed Marek's disease-associated lymphomas, suggesting that Meq is involved in lymphocyte transformation. A revertant virus that restored the expression of the meq gene, showed properties similar to those of the parental virus, confirming that Meq is involved in transformation but not in lytic replication in chickens.

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Selective Cleavage of Human DNA: RecA-Assisted Restriction Endonuclease (RARE) Cleavage

Cited by 201 publications

References 19 publications

RAD51-associated Protein 1 (RAD51AP1) Interacts with the Meiotic Recombinase DMC1 through a Conserved Motif

RAD51-associated Protein 1 (RAD51AP1) Interacts with the Meiotic Recombinase DMC1 through a Conserved Motif

Ordered intracellular RecA–DNA assemblies: A potential site of in vivo RecA-mediated activities

Marek's disease virus-encoded Meq gene is involved in transformation of lymphocytes but is dispensable for replication

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