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DNA Primase (0):
Recent article
DNA Primase
Journal of Hepatology (54)3 2011
Abstract
We examined the molecular basis of iron overload in a family of Vietnamese origin, characterized the molecular and cellular defect, and correlated it with the clinical and pathological phenotype.
We analyzed the ferroportin gene by DNA sequencing. The molecular characterization was p...
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PMID: 21094556
PDF is available here.
PDF is available here.
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Journal of Biological Chemistry (285)43 2010
Abstract
We recently determined the structure of a previously unrecognized pol-prim domain (p68N) that docks on Tag, identified the p68N surface that contacts Tag, and demonstrated its vital role in primosome function. Here, we identify the p68N-docking site on Tag by using structure-guided mutagenesis of th...
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PMID: 20685648
PDF is available here.
PDF is available here.
Abstract
We describe a link between transposition of IS608 and ISDra2, both members of the IS200/IS605 family, which uses obligatory single-stranded DNA intermediates, and the host replication fork. Replication direction through the IS plays a crucial role in excision: activity is maximal when the "top" IS s...
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PMID: 20691900
PDF is available here.
PDF is available here.
Journal of Virology (84)13 2010
Abstract
We demonstrate that Pol-prim plays a major role in the species specificity of BKV DNA replication. Both large subunits p180 and p68 of the enzyme complex have central functions in modulating the host specificity. Recently, an inhibitory activity of BKV DNA replication was described (C. Mahon, B. Lia...
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PMID: 20392840
PDF is available here.
PDF is available here.
Abstract
Previous studies suggested that helicase-primase inhibitor (HPI) resistance mutations can be selected at relatively high frequency from some isolates of herpes simplex virus type 1 (HSV-1). An intentional mismatch primer (IMP) PCR was developed to detect three known HPI resistance mu...
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PMID: 20453068
PDF is available here.
PDF is available here.
Abstract
We have identified 56-kDa DNA primases defective in primer delivery by screening for their ability to support growth of T7Delta4 phage in the presence of this suppressor. Trp69 is critical for primer delivery. Replacement of Trp69 with lysine in either the 56- or 63-kDa gp4 results in defective prim...
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PMID: 20439755
PDF is available here.
PDF is available here.
Abstract
We show, using the T7 replication proteins, that RNA primers are made 'on the fly' during ongoing DNA synthesis and that the leading-strand T7 replisome does not pause during primer synthesis, contrary to previous reports. Instead, the leading-strand polymerase remains limited by the speed of the he...
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PMID: 19924126
PDF is available here.
PDF is available here.
Biochemistry (Washington) (48)31 2009
Abstract
We report unique mechanistic characteristics of recombinant DNA primase from Bacillus anthracis. The mechanism of action of B. anthracis DNA primase (DnaG(BA)) may be described in several distinct steps as follows. Its mechanism of action is initiated when it binds to single-stranded DNA (ssDNA) in...
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PMID: 19583259
PDF is available here.
PDF is available here.
Parasitology (136)7 2009
Abstract
We report the presence of a prex orthologue in T. gondii. The protein is predicted to have a bi-partite apicoplast targeting sequence similar to that demonstrated on the PfPREX polypeptide, capable of delivering marker proteins to the apicoplast. Unlike the P. falciparum gene that is devoid of intro...
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PMID: 19402939
PDF is available here.
PDF is available here.
Abstract
We previously reported on the purification and characterization of a functional multi-protein DNA replication complex (the DNA synthesome) from human cells and tissues. The synthesome is fully competent to carry-out all phases of the DNA replication process in vitro. In this study, DNA primase, a co...
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PMID: 19204933
PDF is available here.
PDF is available here.
Abstract
Vaccinia virus encodes a 90-kDa protein conserved in all poxviruses, with DNA primase and nucleoside triphosphatase activities. DNA primase products, synthesized with a single stranded varphiX174 DNA template, were resolved as dinucleotides and long RNAs on denaturing polyacrylamide and agarose gels...
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PMID: 19007959
PDF is available here.
PDF is available here.
Virus Genes (37)2 2008
Abstract
We have mapped putative motifs throughout these HCMV replication genes. Interesting motifs identified include casein kinase-2 (CKII) phosphorylation sites, a microbodies signal motif in UL57, and an ATP binding site in the putative UL105 helicase. Our investigations have also elucidated motif-rich r...
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PMID: 18618235
PDF is available here.
PDF is available here.
Nucleic Acids Research (36)16 2008
Abstract
We investigated whether adaptations for growth at high temperatures would alter a key functional component of the replication machinery, specifically DnaG primase. Although the structure of bacterial primases is conserved, the trinucleotide initiation specificity for A. aeolicus was hypothesized to...
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PMID: 18684998
PDF is available here.
PDF is available here.
Biochemistry (Washington) (47)34 2008
Abstract
We analyzed the interaction of nucleoside triphosphates (NTPs) containing modified sugars to develop a better understanding of how DNA primase from herpes simplex virus I catalyzes primer synthesis. During the NTP binding reaction, primase tolerated a large number of modifications to the sugar ring....
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PMID: 18672908
PDF is available here.
PDF is available here.
Abstract
We present a series of observations that suggests that mitochondrial RNA polymerase (POLRMT) can act as lagging-strand primase in mammalian cells. POLRMT is highly processive on double-stranded DNA, but synthesizes RNA primers with a length of 25 to 75 nt on a single-stranded template. The short RNA...
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PMID: 18685103
PDF is available here.
PDF is available here.
Abstract
We discuss the mechanism of primer synthesis by RNA polymerase and compare it to mechanisms used by primases of both types as well as to the mechanisms used by RNA polymerase during transcription....
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PMID: 18373065
PDF is available here.
PDF is available here.
Nucleic Acids Research (36)13 2008
Abstract
We report the development of a primase-based Whole Genome Amplification (pWGA) method, which utilizes gp4 primase to synthesize primers, eliminating the requirement of adding synthetic primers. Typical yield of pWGA from 1 ng to 10 ng of human genomic DNA input is in the microgram range, reaching ov...
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PMID: 18559358
PDF is available here.
PDF is available here.
Abstract
The DNA helicase encoded by bacteriophage T7 consists of six identical subunits that form a ring through which the DNA passes. Binding of ssDNA is a prior step to translocation and unwinding of DNA by the helicase. Arg-493 is located at a conserved structural motif within the interior cavity of the...
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PMID: 18574147
PDF is available here.
PDF is available here.
Molecular Microbiology (68)6 2008
Abstract
The study of primases from model organisms such as Escherichia coli, phage T7 and phage T4 has demonstrated the essential nature of primase function, which is to generate de novo RNA polymers to prime DNA polymerase. However, little is known about the function of primases from other eubacteria. Thei...
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PMID: 18452509
PDF is available here.
PDF is available here.
Abstract
By measuring the relative inhibitory concentrations required to overcome particular mutations in the helicase and primase proteins, evidence was obtained that BAY 57-1293 interacts with both components of the helicase-primase complex to achieve maximum potency, whereas for BILS 22BS, this may not be...
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PMID: 18299638
PDF is available here.
PDF is available here.
Molecular Microbiology (68)2 2008
Abstract
We have targeted 10 residues from this network. To investigate the functional contributions of these residues to the primase, ATPase and helicase activities of the primosome, we have used site-directed mutagenesis and in vitro functional assays. Five of these residues (E464, H494, R495, Y548 and R55...
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PMID: 18366438
PDF is available here.
PDF is available here.
Journal of Molecular Biology (377)3 2008
Abstract
We characterized seven different AdPEO-causing mutations in the linker region of TWINKLE and we identified distinct molecular phenotypes. For some mutations, protein hexamerization and DNA helicase activity are completely abolished whereas others display more subtle effects. To better understand the...
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PMID: 18279890
PDF is available here.
PDF is available here.
Journal of Virology (82)5 2008
Abstract
We constructed a DeltaUL52 mutant as well as a complementing cell line. We found that replication of viral DNA was not impaired in noncomplementing cells infected with the DeltaUL52 virus, but viral concatemers remained uncleaved. Since the subnuclear localization of the known cleavage-packaging pro...
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PMID: 18077717
PDF is available here.
PDF is available here.
Abstract
BAY 57-1293 may be more effective than valacyclovir, without the cytotoxicity or potential healing retardation seen with trifluridine. Oral BAY 57-1293 may be a substitute for eye drops as an effective treatment for herpetic keratitis and might be useful in treating stromal keratitis and iritis, as...
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PMID: 18201137
PDF is available here.
PDF is available here.
Oncology Reports (19)2 2008
Abstract
We found that in HeLa cells, p-tyrosyl gallate can effectively induce cell cycle arrest during S phase and inhibited in vitro simian virus (SV40 DNA) replication. In addition, p-tyrosyl gallate can inhibit three important functional replication proteins (topoisomerase I, RPA and pol alpha-primase),...
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PMID: 18202803
PDF is available here.
PDF is available here.
Oncology Reports (19)2 2008
Abstract
We found that polyamines inhibited DNA replication, but it is not clear at which stage this occurs. Spermidine inhibited the DNA cleavage by topoisomerase I at 8.0 mM, but stimulated its activity at 1.0 mM. Spermine also inhibited its activity at 4.0 mM, but stimulated at 1.0 mM. The ssDNA binding a...
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PMID: 18202804
PDF is available here.
PDF is available here.
Abstract
Abstract
We report the full-length crystal structure of G40P, a DnaB family helicase. The hexamer complex reveals an unusual architectural feature and a new type of assembly mechanism. The hexamer has two tiers: a three-fold symmetric N-terminal tier and a six-fold symmetric C-terminal tier. Monomers with tw...
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PMID: 18157148
PDF is available here.
PDF is available here.
Abstract
We have made a number of interesting observations that may be relevant to their clinical use. This article draws attention to the major observations on HPI resistance reported by others and to our own recently published observations that have extended this expanding area of antiviral research....
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PMID: 18610552
PDF is available here.
PDF is available here.
Journal of Molecular Biology (374)5 2007
Abstract
We report on the characterization of the DNA primase complex of the hyperthermophilic archaeon Pyrococcus abyssi (Pab). The Pab DNA primase complex is composed of the proteins Pabp41 and Pabp46, which show sequence similarities to the p49 and p58 subunits, respectively, of the eukaryotic polymerase...
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PMID: 17991487
PDF is available here.
PDF is available here.
Abstract
We report that individual conserved aspartic acid residues in the predicted primase active site were required for in vivo complementation of infectious virus formation as well as genome and plasmid replication. Furthermore, purified recombinant D5 protein synthesized oligoribonucleotides in vitro. I...
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PMID: 18000036
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (282)46 2007
Abstract
Our results offer new horizons in which to investigate the function of high potential [4Fe-4S] clusters in DNA-processing machinery....
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PMID: 17893144
PDF is available here.
PDF is available here.
Abstract
A taxonomic reclassification of the neonatal pathogen Enterobacter sakazakii to consist of five species within a new genus, "Cronobacter," has recently been proposed. The correct identification of these organisms is important to clinicians. Therefore, using 312 Enterobacteriaceae, including 210 "Cro...
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PMID: 17881547
PDF is available here.
PDF is available here.
Abstract
The complex between the DnaB helicase and the DnaG primase unwinds duplex DNA at the eubacterial replication fork and synthesizes the Okazaki RNA primers. The crystal structures of hexameric DnaB and its complex with the helicase binding domain (HBD) of DnaG reveal that within the hexamer the two do...
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PMID: 17947583
PDF is available here.
PDF is available here.
Microbiology (153)Pt 10 2007
Abstract
Invariant and highly conserved amino acids within a primase consensus sequence were targeted by site-specific mutations within the putative primase of Streptococcus thermophilus phage kappa3. PCR products containing the desired mutation(s) within putative ATPase/helicase and/or oligomerization domai...
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PMID: 17906129
PDF is available here.
PDF is available here.
FEBS Letters (581)21 2007
Abstract
Replication protein A (RPA) is a stable heterotrimeric complex consisting of p70, p32 and p14 subunits. The protein plays a crucial role in SV40 minichromosome replication. Peptides of p70 representing interaction sites for the smaller two subunits, DNA as well as the viral initiator protein large T-...
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PMID: 17673209
PDF is available here.
PDF is available here.
Abstract
We show that, in haploid budding yeast, Rad51-dependent BIR induced by HO endonuclease requires the lagging strand DNA Polalpha-primase complex as well as Poldelta to initiate new DNA synthesis. Polepsilon is not required for the initial primer extension step of BIR but is required to complete 30 kb...
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PMID: 17671506
PDF is available here.
PDF is available here.
Journal of Virology (81)16 2007
Abstract
We constructed an HSV-1 UL52 L986F mutation and found that it can complement a UL52 null virus more efficiently than the wild type (WT). We thus predicted that the UL5/8/52 complex containing the L986F mutation might possess increased primase activity; however, it exhibited only 25% of the WT level...
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PMID: 17553899
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (282)29 2007
Abstract
Our presteady state kinetic studies of helicases from two classes, the ring-shaped T7 helicase and two forms of non-ring-shaped hepatitis C virus (HCV) helicase, show that both the unwinding rate and processivity depend on the sequence and decrease as the nucleic acid stability increases. The DNA un...
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PMID: 17504766
PDF is available here.
PDF is available here.
Abstract
While plasmids are very commonly associated with the majority of the lactic acid bacteria, they are only very rarely associated with Lactobacillus delbrueckii, with only four characterized to date. In this study, the complete sequence of a native plasmid, pDOJ1, from a strain of Lactobacillus delbru...
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PMID: 17526779
PDF is available here.
PDF is available here.
Abstract
We have developed a rapid and effective pool-based screen, termed FASTDXL (focused array screening technique for disulfide X-linking), to produce and identify disulfide-stabilized protein-nucleic acid assemblies. A significant strength of FASTDXL is that it can take advantage of prior structural kno...
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PMID: 17637338
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (282)27 2007
Abstract
We show here that PriA binds to DNA in a manner either dependent on or independent of 3' terminus recognition. The former mode of binding requires the 3' terminus binding pocket present at the N-terminal half of the 181-residue DNA binding domain and exhibits specific bipartite interaction on the te...
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PMID: 17483094
PDF is available here.
PDF is available here.
Biochimica et Biophysica Acta (1774)6 2007
Abstract
The ring-shaped helicases represent one important group of helicases that can translocate along single-stranded (ss) DNA and unwinding double-stranded (ds) DNA by using the energy derived from NTP binding and hydrolysis. Despite intensive studies, the mechanism by which the ring-shaped helicase tran...
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PMID: 17499029
PDF is available here.
PDF is available here.
Abstract
We applied virtual screening to discover primase inhibitors, and subsequently several commercially available analogs of these initial hits showed potent primase inhibition and in vitro antibacterial activity. This work provides a 3D pharmacophore for primase ligands, SAR trends, and leads that can b...
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PMID: 17350255
PDF is available here.
PDF is available here.
Abstract
Abstract
A novel series of imidazolylpyrimidines were found to possess inhibitory activity against the human CMV UL70 primase. Extensive SAR studies on an HTS lead led to potent, orally bioavailable compounds with anti-CMV IC(50) values of 150 nM in both viral yield and viral DNA replication assays and with...
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PMID: 17329103
PDF is available here.
PDF is available here.
Journal of Virology (81)8 2007
Abstract
The effect of Epstein-Barr virus (EBV) SM protein on EBV gene expression was examined using a recombinant EBV strain with the SM gene deleted and DNA microarrays representing all known EBV coding regions. Induction of lytic EBV replication in the absence of SM led to expression of approximately 40%...
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PMID: 17287267
PDF is available here.
PDF is available here.
Abstract
We characterized a mechanism that regulates replication elongation in the bacterium Bacillus subtilis. Replication elongation was inhibited within minutes after amino acid starvation, regardless of where the replication forks were located on the chromosome. We found that small nucleotides ppGpp and...
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PMID: 17350574
PDF is available here.
PDF is available here.
Abstract
The bacterial signaling molecules ppGpp and pppGpp regulate transcription initiation in response to starvation by altering RNA polymerase activity. In this issue, Wang et al. (2007) show that (p)ppGpp also inhibits DNA replication elongation by interfering with DNA primase activity. Halting replicat...
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PMID: 17350566
PDF is available here.
PDF is available here.
Molecular Microbiology (63)6 2007
Abstract
We have employed a 'domain swapping' approach to replace p17 with its structural homologue p16 to create chimeras. p33 alone hydrolyses ATP but exhibits no helicase activity. Fusing p16 (p16-p33) or DnaG (G-p33) to the N-terminus of p33 produced chimeras with partially restored helicase activities....
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PMID: 17367384
PDF is available here.
PDF is available here.
Molecular Microbiology (63)3 2007
Abstract
We show that the small subunit of the clamp loader replication factor C (RFC) of S. solfataricus interacted with both the catalytic and non-catalytic subunits of the primase by yeast two-hybrid and co-immunoprecipitation assays. Further, the primase-RFC interaction was also identified in the cell ex...
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PMID: 17181784
PDF is available here.
PDF is available here.
EMBO Reports (8)1 2007
Abstract
We report the biochemical characterization of the human GINS complex (hGINS). The four hGINS subunits were coexpressed in Escherichia coli in a highly soluble form and purified as a complex. hGINS was shown to interact directly with the heterodimeric human DNA primase, by using either surface plasmo...
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PMID: 17170760
PDF is available here.
PDF is available here.
Abstract
We confirmed that BAY 57-1293 is highly active against HSV-1 and superior to ACV when tested in Vero cells. Notably, drug resistance was detected in laboratory working stocks in two different strains of HSV at 10(-4) to 10(-5) and there was evidence that the resistant variants were present in the vi...
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PMID: 17354648
PDF is available here.
PDF is available here.
Abstract
The efficacy of BAY 57-1293, a novel non-nucleosidic inhibitor of herpes simplex virus 1 and 2 (HSV-1 and HSV-2), bovine herpesvirus and pseudorabies virus, was studied in the guinea pig model of genital herpes in comparison with the licensed drug valaciclovir (Valtrex). Early therapy with BAY 57-12...
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PMID: 17354650
PDF is available here.
PDF is available here.
Nucleic Acids Research (35)4 2007
Abstract
We set out to produce GpppA- and (7Me)GpppA-capped RNA oligonucleotides for non-radioactive mRNA cap methyltransferase assays and, in perspective, for studies of enzyme specificity in relation to substrate length as well as for co-crystallization studies. This study reports the use of a bacteriophag...
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PMID: 17259217
PDF is available here.
PDF is available here.
Archives of Virology (152)8 2007
Abstract
We present evidence that single mutations close to a predicted functional domain of an essential HSV-1 replication enzyme (helicase) are associated with drug resistance and virus growth characteristics....
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PMID: 17404685
PDF is available here.
PDF is available here.
Archives of Virology (152)8 2007
Abstract
Little is known about the bacteriophage proteins expressed immediately after infection of the host cell. Most of these early proteins are probably involved in bacteriophage-host interactions redirecting the bacterial metabolism to phage production. Interaction analysis of the first 16 phiKMV gene pr...
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PMID: 17534690
PDF is available here.
PDF is available here.
Nucleic Acids Research (35)17 2007
Abstract
Primases are specialized DNA-dependent RNA polymerases that synthesize a short oligoribonucleotide complementary to single-stranded template DNA. In the context of cellular DNA replication, primases are indispensable since DNA polymerases are not able to start DNA polymerization de novo. The primase...
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PMID: 17709343
PDF is available here.
PDF is available here.
Abstract
We speculate that the replication mechanism for the TBB phytoplasma eDNA involves the dnaG gene instead of the rep gene. The Ca. P. australiense eDNA (3773 bp) was shown to be circular and contained four open reading frames. The rep gene was encoded on ORF 1 and had homology to both plasmid (pLS1) a...
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PMID: 16879868
PDF is available here.
PDF is available here.
Molecular Microbiology (62)4 2006
Abstract
We characterize the exosome of Sulfolobus solfataricus. Two exosome fractions can be discriminated by density gradient centrifugation. We show that the Cdc48 protein is associated with the exosome from the 30S-50S fraction but not with the exosome of the 11.3S fraction. While only some complexes con...
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PMID: 17078816
PDF is available here.
PDF is available here.