Traditionally, recombination reactions promoted by RecA-like proteins initiate by forming a nucleoprotein filament on a single-stranded DNA (ssDNA), which then pairs with homologous double-stranded DNA (dsDNA). In this paper, we describe a novel pairing process that occurs in an unconventional manner: RecA protein polymerizes along dsDNA to form an active nucleoprotein filament that can pair and exchange strands with homologous ssDNA. Our results demonstrate that this inverse reaction is a unique, highly efficient DNA strand exchange reaction that is not due to redistribution of RecA protein from dsDNA to the homologous ssDNA partner. Finally, we demonstrate that the RecA protein-dsDNA filament can also pair and promote strand exchange with ssRNA. This inverse RNA strand exchange reaction is likely responsible for R-loop formation that is required for recombination-dependent DNA replication.
1. RaddingCM 1981 Recombination activities of E. coli recA protein. Cell 25 3 4. 2. LusettiSLCoxMM 2002 The bacterial RecA protein and the recombinational DNA repair of stalled replication forks. Annu Rev Biochem 71 71 100. 3. CoxMM 2007 Regulation of bacterial RecA protein function. Crit Rev Biochem Mol Biol 42 41 63. 4. CoxMM 2007 Motoring along with the bacterial RecA protein. Nat Rev Mol Cell Biol 8 127 138. 5. TamasIKlassonLCanbackBNaslundAKErikssonAS 2002 50 million years of genomic stasis in endosymbiotic bacteria. Science 296 2376 2379. 6. SeitzEMBrockmanJPSandlerSJClarkAJKowalczykowskiSC 1998 RadA protein is an archaeal RecA protein homolog that catalyzes DNA strand exchange. Genes Dev 12 1248 1253. 7. ShinoharaAOgawaHOgawaT 1992 Rad51 protein involved in repair and recombination in S. cerevisiae is a RecA-like protein. Cell 69 457 470. 8. VamvakasSVockEHLutzWK 1997 On the role of DNA double-strand breaks in toxicity and carcinogenesis. Crit Rev Toxicol 27 155 174. 9. KhannaKKJacksonSP ...
TY - JOUR. T1 - Negative co-dominant inhibition of recA protein function. T2 - Biochemical properties of the recA1, recA13 and recA56 proteins and the effect of recA56 protein on the activities of the wild-type recA protein function in vitro. AU - Lauder, Scott D.. AU - Kowalczykowski, Stephen C.. PY - 1993. Y1 - 1993. N2 - We have investigated the biochemical properties of several Escherichia coli mutant recA proteins that display a null phenotype. These are the recA1, recA13 and recA56 proteins, each of which carries a single missense mutation. These proteins all share a common defect which is the inability to adopt the high affinity DNA binding state normally elicited by the nucleotide cofactor ATP. Consequently, other than the ability to bind ssDNA, they possess none of the in vitro enzymatic activities of recA protein. However, each protein has characteristics that are unique, leading to the conclusion that the observed mutant phenotypes arise through fundamentally different mechanisms. ...
The effect of the cellular level of RecA protein on the ability of E. coli K12 bacteria to (i) survive UV-irradiation (ii) promote UV-reactivation of UV-damaged phage lambda (iii) induce prophage lambda was determined in bacterial mutants with discrete increasing levels of RecA protein. The various levels of RecA protein were obtained by combining lexA and recA alleles. Except for the double mutant lexA3 recAo98, whose repair ability was 25% less than that observed in wild type bacteria, bacterial survival was proportional to the level of RecA protein measured after 90 min of incubation. In lexA3 recAo98 bacteria, RecA protein, at a constitutive high basal level, failed to compensate totally for the lack of LexA repressor cleavage; UV-reactivation of UV-damaged phage lambda was not restored; yet, prophage lambda was induced with 35% efficiency. Efficient UV-induction of prophage lambda is linked to the induction of lexA-controlled host processes that repair the UV-damaged prophage.
The RecA730 protein is able to nucleate itself onto single stranded DNA and suppress the recombinational defect of a recF mutant. By determining cell survival after UV and gamma irradiation and conjugational recombination proficiency, we concluded that this mutant could suppress the deficiency of cells lacking RecA loading functions from both pathways (RecBCD, RecFOR), as well as the helicase deficiency in the recB1080 recQ mutant. It could not suppress the absence of exonuclease functions from both pathways (RecBCD, RecJ). RecA filament formation and its role in recombination and DNA repair depends on the RecA proteins ability to bind and hydrolyze ATP. The addition of the recA730 mutation in the recA2201 mutant, which has the phenotype of the recA null mutant since it is not able to hydrolyze ATP, results in a conformational change that enables the double mutant recA730, 2201 to induce the SOS response. By determining the basal level of the SOS response and the level after introduction of a ...
Eukaryotic cells encode two homologs of Escherichia coli RecA protein, Rad51 and Dmc1, which are required for meiotic recombination. Rad51, like E.coli RecA, forms helical nucleoprotein filaments that promote joint molecule and heteroduplex DNA formation. Electron microscopy reveals that the human meiosis-specific recombinase Dmc1 forms ring structures that bind single-stranded (ss) and double-stranded (ds) DNA. The protein binds preferentially to ssDNA tails and gaps in duplex DNA. hDmc1-ssDNA complexes exhibit an irregular, often compacted structure, and promote strand-transfer reactions with homologous duplex DNA. hDmc1 binds duplex DNA with reduced affinity to form nucleoprotein complexes. In contrast to helical RecA/Rad51 filaments, however, Dmc1 filaments are composed of a linear array of stacked protein rings. Consistent with the requirement for two recombinases in meiotic recombination, hDmc1 interacts directly with hRad51.. ...
The Escherichia coli protein RecA catalyzes the strand exchange reaction used in DNA repair and genetic recombination. RecA is also a target for inhibiting microbial antibiotic resistance, understanding cancer propagation, and characterizing neurodegenerative disorders. Therefore, understanding factors that affect RecA structure and stability is broadly applicable to many fields. Previous studies in our lab have shown buffer-specific changes in RecA stability and unfolding transitions. These studies suggest only minimal buffer-dependent changes in nucleotide binding and secondary structure but do not explain the significant differences in RecA stability and unfolding profiles. Here we have employed various biochemical and spectroscopic techniques to further characterize RecA both structurally and functionally in four common buffers: Tris, MES, HEPES, and Phosphate. Activity assays reveal RecA activity may be slightly decreased in HEPES and Phosphate buffers as compared to Tris and MES. Circular
Can catalyze the hydrolysis of ATP in the presence of single-stranded DNA, the ATP-dependent uptake of single-stranded DNA by duplex DNA, and the ATP-dependent hybridization of homologous single-stranded DNAs. It interacts with LexA causing its activation and leading to its autocatalytic cleavage.
Can catalyze the hydrolysis of ATP in the presence of single-stranded DNA, the ATP-dependent uptake of single-stranded DNA by duplex DNA, and the ATP-dependent hybridization of homologous single-stranded DNAs. It interacts with LexA causing its activation and leading to its autocatalytic cleavage.
The RAD51 gene of Saccharomyces cerevisiae is required both for recombination and for the repair of DNA damage caused by X rays. Here we report the sequence and transcriptional regulation of this gene. The RAD51 protein shares significant homology (approximately 50%) over a 70-amino-acid with the RAD57 protein (J.A. Kans and R.K. Mortimer, Gene 105:139-140, 1991), the product of another yeast recombinational repair gene, and also moderate (approximately 27%), but potentially significant, homology with the bacterial RecA protein. The homologies cover a region that encodes a putative nucleotide binding site of the RAD51 protein. Sequences upstream of the coding region for RAD51 protein share homology with the damage response sequence element of RAD54, an upstream activating sequence required for damage regulation of the RAD54 transcript, and also contain two sites for restriction enzyme MluI; the presence of MluI restriction sites has been associated with cell cycle regulation. A 1.6-kb transcript ...
Protein filaments in Alzheimers disease. Coloured Transmission Electron Micrograph (TEM) of beta protein filaments which occur in brain tissue in Alzheimers disease. The filaments here have been purified in the laboratory. Alzheimers disease is a form of senile dementia. The brain is affected by degenerative changes in frontal and temporal lobes, and brain tissue shrinks in size. Senile plaques are produced which consist, as seen here, of tangled masses of filaments and granules which are often centred around an area of amyloid. Masses of thickened filaments also form in the cytoplasm of nerve cells, called neurofibrillary tangles. Magnification: x7,000 at 35mm size. - Stock Image M108/0321
BRCC5, hRAD51, HRAD51, HsRad51, HsRAD51, HsT16930, RAD51 (S. cerevisiae) homolog (E coli RecA homolog), RAD51 homolog (RecA homolog, E. coli) (S. cerevisiae), RAD51 homolog A, RAD51ABRCA1/BRCA2-containing complex, subunit 5, RecA, E. coli, homolog of, RECADNA repair protein RAD51 homolog 1, RecA-like protein, recombination protein ...
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RAD51 recombinase, the eukaryotic homolog of RecA, is the core component in homologous recombination (HR). It performs fundamental roles such as homologous pairing and strand exchange in repairing DNA double strand breaks [1, 2]. Rad51 binds both ssDNA/dsDNA [3-5] and forms right-handed helical nucleoprotein filaments in which DNA is extended 1.5 times that of B form DNA [4, 6-8]. Such extended conformation of DNA in ssDNA-protein filaments is instrumental in facilitating homology search and strand exchange in a three stranded pairing system [8-10]. Even though the equivalent scenario of extended DNA configuration is demonstrable in dsDNA-protein filament, its relevance in HR is not fully understood. However, based on E. coli RecA system, it is surmised that unwinding associated with dsDNA-protein filament is similarly important in four-stranded pairing and strand exchange processes [11]. The first and also most recent report that describes four-stranded exchange reaction catalyzed by RAD51 ...
A RecA-single-stranded DNA (RecA-ssDNA) filament searches a genome for sequence homology by rapidly binding and unbinding double-stranded DNA (dsDNA) until homology is found. We demonstrate that pulling on the opposite termini (3′ and 5′) of one of the two DNA strands in a dsDNA molecule stabilizes the normally unstable binding of that dsDNA to non-homologous RecA-ssDNA filaments, whereas pulling on the two 3′, the two 5′, or all four termini does not. We propose that the outgoing strand in the dsDNA is extended by strong DNA-protein contacts, whereas the complementary strand is extended by the tension on the base pairs that connect the complementary strand to the outgoing strand. The stress resulting from different levels of tension on its constitutive strands causes rapid dsDNA unbinding unless sufficient homology is present ...
S-DNA (stretched DNA) is an elongated base-paired DNA conformation under high tension. Because the RecA/Rad51 family DNA recombinases form helical filaments on DNA and mediate the formation of the DNA triplex (D-loop), in which the DNA is stretched, and because the extension of these nucleoprotein filaments is similar to the extension of S-DNA, S-DNA has long been hypothesized as a possible state ...
For the first time, scientists have created from scratch self-assembling protein filaments built from identical protein subunits that snap together spontaneously to form long, helical, thread-like structures. Protein filaments are important to our bodies. In nature, protein filaments play a key role in cell biology.
The SOS response in Eubacteria is a global response to DNA damage and its activation is increasingly associated with the movement of mobile genetic elements. The temperate phage GIL01 is induced into lytic growth using the hosts SOS response to genomic stress. LexA, the SOS transcription factor, represses bacteriophage transcription by binding to a set of SOS boxes in the lysogenic promoter P1. However, LexA is unable to efficiently repress GIL01 transcription unless the small phage-encoded protein gp7 is also present. We found that gp7 forms a stable complex with LexA that enhances LexA binding to phage and cellular SOS sites and interferes with RecA-mediated auto-cleavage of LexA, the key step in the initiation of the SOS response. Gp7 did not bind DNA, alone or when complexed with LexA. Our findings suggest that gp7 induces a LexA conformation that favors DNA binding but disfavors LexA auto-cleavage, thereby altering the dynamics of the cellular SOS response. This is the first account of an ...
Filaments are also highly dynamic in nature and far from a static structure that acts as a stable "scaffold" for a cell.[citation needed] Some phenomena that profile a protein filaments dynamics are:[citation needed] ...
In Escherichia coli, the SOS response is induced upon DNA damage and results in the enhanced expression of a set of genes involved in DNA repair and other functions. The initial step, self-cleavage of the LexA repressor, is promoted by the RecA protein which is activated upon binding to single-stranded DNA. In this work, induction of the SOS response by the addition of mitomycin C was found to be prevented by overexpression of the dinI gene. dinI is an SOS gene which maps at 24.6 min of the E.coli chromosome and encodes a small protein of 81 amino acids. Immunoblotting analysis with anti-LexA antibodies revealed that LexA did not undergo cleavage in dinI-overexpressed cells after UV irradiation. In addition, the RecA-dependent conversion of UmuD to UmuD (the active form for mutagenesis) was also inhibited in dinI-overexpressed cells. Conversely, a dinI-deficient mutant showed a slightly faster and more extensive processing of UmuD and hence higher mutability than the wild-type. Finally, we ...
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InterPro provides functional analysis of proteins by classifying them into families and predicting domains and important sites. We combine protein signatures from a number of member databases into a single searchable resource, capitalising on their individual strengths to produce a powerful integrated database and diagnostic tool.
InterPro provides functional analysis of proteins by classifying them into families and predicting domains and important sites. We combine protein signatures from a number of member databases into a single searchable resource, capitalising on their individual strengths to produce a powerful integrated database and diagnostic tool.
The RAD 51 protein, a eukaryotic homologue of Escherichia coli RecA, plays a significant role in both mitotic and meiotic homologous recombination. Here, we demonstrate that short-term silencing of th
Product from Supplier BBridge, Catalog number 61-001, Product anti-E.coli LexA antibody, rabbit serum Antibodies DNA Repair_Recombination - Gentaur molecular products
Biegeleisen, K. (2002) Topologically Non-Linked Circular Duplex DNA. |i|Bulletin of Mathematical Biology|/i|, 64, 589- 609. |br /| |a href=http://dx.doi.org/10.1006/bulm.2002.0288 target=_blank|http://dx.doi.org/10.1006/bulm.2002.0288|/a|
Intragenic recombination between the single complete pilin gene (expression locus) and multiple, distinct, partial pilin gene copies (silent, storage loci) is thought to account for the generation of pilus antigenic diversity and piliation phase (on-off) changes exhibited by Neisseria gonorrhoeae. The mechanisms operating in the genomic rearrangements associated with these forms of pilus variation were investigated through the study of isogenic strains of gonococci bearing either wild-type or altered recA alleles. Examination of the rates of pilus phase variation and the genetic basis for changes in piliation status displayed by these strains show that recA mediated homologous recombination is required for these high frequency events and confirm that the nonpiliated state results from mutations in the expressed pilin gene. In a strain that is deficient in recA mediated homologous recombination, pilus phase variation occurs at a 100-1000-fold reduced rate and results predominantly from one class ...
The protein encoded by this gene is essential for meiotic homologous recombination. Genetic recombination in meiosis plays an important role in generating diversity of genetic information. The product of this gene is structurally and evolutionary related to the products of the yeast RAD51 and E. coli RecA genes. Alternative splice variants of this gene have been described but their full-length nature has not been determined.
SWISS-MODEL Template Library (SMTL) entry for 4po9.1. Mycobacterium tuberculosis RecA glycerol bound low temperature structure IIC-BR
Gruber AJ, Erdem AL, Sabat G, Karata K, Jaszczur MM, Vo DD, Olsen TM, Woodgate R, Goodman MF, Cox MM. A RecA protein surface required for activation of DNA polymerase V. PLoS Genet 2015; 11(3):e1005066. Robinson A, McDonald JP, Caldas VE, Patel M, Wood EA
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The cytoskeleton is a complex network of protein filaments extending throughout the cytoplasm that is involved in a range of cell processes. Of the three main
HOMOLOGOUS recombination is required for the faithful repair of DNA double-strand breaks (DSBs) that arise during normal cellular processes or from exposure of cells to DNA-damaging agents. Central to the process of homologous recombination is the Rad51 protein, which facilitates synapsis and strand invasion into homologous duplex DNA (San Filippo et al. 2008). Rad51 belongs to the RecA family of homologous pairing proteins (Aboussekhra et al. 1992; Basile et al. 1992; Shinohara et al. 1992). Yeast and humans have two RecA homologs: Rad51 and the meiosis-specific Dmc1 (Bishop et al. 1992; San Filippo et al. 2008). In addition, the Saccharomyces cerevisiae RAD55 and RAD57 genes encode proteins with sequence similarity to RecA and Rad51 and are considered to be Rad51 paralogs (Kans and Mortimer 1991; Lovett 1994). Mutation of RAD51, RAD55, or RAD57 confers sensitivity of ionizing radiation (IR) and defects in mitotic and meiotic recombination, indicating that their functions are not redundant ...
Spatial regulation is often encountered as a component of multi-tiered regulatory systems in eukaryotes, where processes are readily segregated by organelle boundaries. Well-characterized examples of spatial regulation are less common in bacteria. Low-fidelity DNA polymerase V (UmuD′2C) is produced in Escherichia coli as part of the bacterial SOS response to DNA damage. Due to the mutagenic potential of this enzyme, pol V activity is controlled by means of an elaborate regulatory system at transcriptional and posttranslational levels. Using single-molecule fluorescence microscopy to visualize UmuC inside living cells in space and time, we now show that pol V is also subject to a novel form of spatial regulation. After an initial delay (~ 45 min) post UV irradiation, UmuC is synthesized, but is not immediately activated. Instead, it is sequestered at the inner cell membrane. The release of UmuC into the cytosol requires the RecA* nucleoprotein filament-mediated cleavage of UmuD→UmuD′. Classic SOS
RecA is important in recombination, DNA repair and repair of replication forks. It functions through the production of a protein-DNA filament. To study the localization of RecA in live Escherichia coli cells, the RecA protein was fused to the green fluorescence protein (GFP). Strains with this gene …
From the above, it is clear that filament architectures and mechanisms of assembly range from the simple isodesmic to extremely complex two‐protein matrix‐associated collaborative filament systems, and examples of such complex collaborative assemblies are probably fairly common as they are important components of large, self‐assembling systems. The addition of a lateral binding partner to all, or a subset of subunits along the filament, either through a neighbouring matrix or indeed another filament creates opportunities for complex and emerging properties of the resulting systems.. The most obvious, but by no means only consequence is that collaborative filaments may be restricted in occurrence to the site of the matrix or scaffold they bind to.. The large number of additional, lateral binding sites sometimes creates large cooperativity effects that enable filaments to bind with extreme affinity to their partner, even if the individual binding energies, per subunit, are rather small. The ...
In the simplest form of three-strand braid, all the hair is initially divided into three sections, which are then simultaneously gathered together near the scalp. In contrast, a French braid starts with three small sections of hair near the crown of the head, which are then braided together toward the nape of the neck, gradually adding more hair to each section as it crosses in from the side into the center of the braid structure. The final result incorporates all of the hair into a smoothly woven pattern over the scalp. If the main mass of hair is initially parted into two or more sections along the scalp that are kept separate from one another, multiple French braids may be created, each in its own section. One unique feature about the French braid is that an individual can braid their own hair without the help of others. The difficulty of braiding can depend on the type of hair the individual has, some styles of hair are easier to braid than others. The length of hair also plays a role in the ...
Very little is known about the role of DNA repair networks in Brucella abortus and its role in pathogenesis. We investigated the roles of RecA protein DNA repair and SOS regulation in B. abortus. While recA mutants in most bacterial species are hyper ...
HMS174(DE3) Competent Cells - Novagen HMS174 strains provide high transformation efficiencies and the recA mutation in a K-12 background. Strain may stabilize certain target genes whose products may cause the loss of the DE3 prophage. - Find MSDS or SDS, a COA, data sheets and more information.
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Lexa 400 Abec 7 Spool found in: DAIWA LEXA 400 HS SPOOL/TENSION BAITCASTER Ceramic Orange Seal ABEC 7 Manufacturer/Model By Series, DAIWA LEXA 400 H..
I tried all three flavors of Recor Pre Action and then went to Soulcycle. Im alive to tell the tale. Read my review to hear my full recap of this pre workout.
Ep 7: Mariah brought her security guard and Reco brought his beef with his former BFF. See the Married to Medicine ladies react to fashion show fight.
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Accessory replicative helicases aid the primary replicative helicase in duplicating protein-bound DNA, especially transcribed DNA. Recombination enzymes also aid genome duplication by facilitating the repair of DNA lesions via strand exchange and also processing of blocked fork DNA to generate structures onto which the replisome can be reloaded. There is significant interplay between accessory helicases and recombination enzymes in both bacteria and lower eukaryotes but how these replication repair systems interact to ensure efficient genome duplication remains unclear. Here, we demonstrate that the DNA content defects of Escherichia coli cells lacking the strand exchange protein RecA are driven primarily by conflicts between replication and transcription, as is the case in cells lacking the accessory helicase Rep. However, in contrast to Rep, neither RecA nor RecBCD, the helicase/exonuclease that loads RecA onto dsDNA ends, is important for maintaining rapid chromosome duplication. Furthermore, RecA
Homologous recombination plays a central role in maintaining genomic DNA as well as cellular tolerance to various DNA-damaging agents (1, 2). Specifically, homologous recombination restores stalled replication at damaged template strands by using the intact sister chromatid as a template for DNA synthesis (3-6) and repairs any double-strand breaks (DSB) that may occur in one of the sister chromatids during replication or as a result of chemotherapeutic treatments with camptothecin, cisplatin, and PARP inhibitor. Individual homologous recombination factors contribute in different ways to various types of homologous recombination (7, 8). In homologous recombination-dependent DSB repair, nucleases initiate homologous recombination by processing DSBs to generate 3′ overhangs, followed by the polymerization of RAD51 recombinase on the overhang (9-11). The resulting RAD51 polymer is responsible for homology search and subsequent strand exchange with intact homologous duplex DNA. RAD51 is required ...
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Streptococcus pneumoniae MmsA protein: RecG-like protein involved in DNA repair and three-strand recombination; isolated from Streptococcus pneumoniae; amino acid sequence in first source; do not confuse with MMSA from Plasmodium
Homologous recombination is a genetic process in which two similar DNA strands exchange material. The main purpose of homologous...
Chris, If you try mitomycin C remember that it is light sensitive. It can be incorporated into growth media (e.g., BHI, etc.) but must be added after autoclaving just as you would an antibiotic such as ampicillin. For best results youll have to determine the proper concentration (i.e., below the MIC) which will vary from strain to strain. I would suggest somewhere between 0.2 to 1 ug/ml as a start. If you decide to go the UV route its really not a problem. You can irradiate a bacterial suspension (just make sure its in saline, not a lot of organics which will absorb UV) in a conventional plastic petri plate with the lid off for the short period of UV exposure they will require. Good luck, R. Goering On Fri, 14 Jan 1994, Christopher Coward wrote: , I want to induce the SOS response in E. coli to investigate mutants deficient , in the exit from the SOS response (ie although they may repair any DNA , damage, they do not resume cell division). Does anyone have any suggestions as , to how to ...