Escherichia coli polynucleotide phosphorylase (PNPase) is generally believed to be involved in mRNA degradation via its 3$\sp\prime$ to 5$\sp\prime$ exoribonuclease activity. However, there are many unexplained observations related to the physiological role of this enzyme. The projects conducted in this thesis are aimed at understanding more about the in vivo function of PNPase. Two approaches were employed to try to achieve this goal. First, a biochemical approach involving the purification of PNPase associated proteins was used. Specifically, the previously documented $\beta$ subunit present in the pentameric form of PNPase was examined as an initial step toward understanding the mechanism and possibly the regulation of PNPase catalysis. Second, a genetic approach was undertaken which focused on characterization of mutant strains lacking PNPase in combination with other known enzymes involved in RNA metabolism, in particular, tRNA nucleotidyltransferase, poly(A) polymerase and RNase PH.^ From the
In this study, the importance of the RPH domains in mediating the characteristic phenotypic changes induced by hPNPaseold-35 is firmly established. We observed that the presence of at least one RPH domain is required for the functional activity of the protein and either of the domains is as potent as the full-length molecule. This contrasts with bacterial PNPase in which mutation in the key residues in either of the RPH domains inhibits catalytic activity (19). However, our result is comparable to chloroplast PNPase in which the first RPH domain alone (comparable to our ΔRPH2 construct) is highly active enzymatically (40). Although the second RPH domain (our ΔRPH1) of the chloroplast has low RNA degradation activity of nonpolyadenylated RNA, it has high activity for polyadenylated RNA (40), which explains the efficiency of the ΔRPH1 construct in degrading human polyadenylated mRNAs. The RPH domains themselves can bind to RNA, and the PNPase domain is also involved in RNA binding, which ...
TY - JOUR. T1 - An essential function for the phosphate-dependent exoribonucleases RNase PH and polynucleotide phosphorylase. AU - Zhou, Zhihua. AU - Deutscher, Murray P.. PY - 1997/7. Y1 - 1997/7. N2 - Escherichia coli cells lacking both polynucleotide phosphorylase (PNPase) and RNase PH, the only known P1-dependent exoribonucleases, were previously shown to grow slowly at 37°C and to display a dramatically reduced level of tRNA(Tryrsu3+) suppressor activity. Here we show that the RNase PH-negative, PNP-negative double-mutant strain actually displays a reversible cold-sensitive phenotype and that tRNA biosynthesis is normal. In contrast, ribosome structure and function are severely affected, particularly at lower temperatures. At 31°C, the amount of 50S subunit is dramatically reduced and 23S rRNA is degraded. Moreover, cells that had been incubated at 42°C immediately cease growing and synthesizing protein upon a shift to 31°C, suggesting that the ribosomes synthesized at the higher ...
RNA import into mammalian mitochondria is considered essential for replication, transcription, and translation of the mitochondrial genome but the pathway(s) and factors that control this import are poorly understood. Previously, we localized polynucleotide phosphorylase (PNPASE), a 3 --, 5 exoribonuclease and poly-A polymerase, in the mitochondrial intermembrane space, a location lacking resident RNAs. Here, we show a new role for PNPASE in regulating the import of nuclear-encoded RNAs into the mitochondrial matrix. PNPASE reduction impaired mitochondrial RNA processing and polycistronic transcripts accumulated. Augmented import of RNase P, 5S rRNA, and MRP RNAs depended on PNPASE expression and PNPASE-imported RNA interactions were identified. PNPASE RNA processing and import activities were separable and a mitochondrial RNA targeting signal was isolated that enabled RNA import in a PNPASE-dependent manner. Combined, these data strongly support an unanticipated role for PNPASE in mediating ...
POLYRIBONUCLEOTIDE NUCLEOTIDYLTRANSFERASE, POLYRIBONUCLEOTIDE NUCLEOTIDYLTRANSFERASE, POLYRIBONUCLEOTIDE NUCLEOTIDYLTRANSFERASE, RNA, 5-R(*UP*AP*AP*CP*UP*UP*UP*GP*GP)-3, RNA, 5-R(*UP*AP*AP*CP*UP*UP*UP*GP*GP)-3, RNA, 5-R(*UP*AP*AP*CP*UP*UP*UP*GP*GP)-3, RNA, 5-R(*UP*AP*AP*CP*UP*UP*UP*GP*GP)-3 ...
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SUMMARY: Polynucleotide phosphorylase has been purified from the cyanobacterium Nostoc sp. MAC. The enzyme requires a divalent cation such as Mg2+, has a pH optimum of 10·5 and catalyses the polymerization of ADP into polynucleotide in a primer-independent reaction at a rate of 2 μmol min-1 (mg protein)-1. It has an apparent native M r of 215000-240000. Non-denaturing polyacrylamide activity gels reveal a heterogeneous pattern of active bands similar to those previously observed with the corresponding enzyme from Micrococcus luteus, while SDS-denaturing activity gels reveal a single band of activity of M r 91000 in both crude extracts and the most highly purified fraction. Activity has also been demonstrated in a cetyltrimethyl-ammonium bromide non-denaturing activity gel. By analogy with other known polynucleotide phosphorylases, the Nostoc enzyme is probably a trimer of the 91000-M r subunit.
Polynucleotide Phosphorylase (PNPase) is a bifunctional enzyme with a phosphorolytic 3 to 5 exoribonuclease activity and a 3-terminal oligonucleotide polymerase activity.[2] That is, it dismantles the RNA chain starting at the 3 end and working toward the 5 end.[1] It also synthesizes long, highly heteropolymeric tails in vivo. It accounts for all of the observed residual polyadenylation in strains of Escherichia coli missing the normal polyadenylation enzyme.[1] Discovered by Marianne Grunberg-Manago working in Severo Ochoas lab in 1955, the RNA-polymerization activity of PNPase was initially believed to be responsible for DNA-dependent synthesis of messenger RNA, a notion that got disproved by the late 1950s.[3][4] It is involved in mRNA processing and degradation in bacteria, plants,[5] and in humans.[6] In humans, the enzyme is encoded by the PNPT1 gene. In its active form, the protein forms a ring structure consisting of three PNPase molecules. Each PNPase molecule consists of two ...
Complete information for PNP gene (Protein Coding), Purine Nucleoside Phosphorylase, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
Typhoid fever occurs almost exclusively in developing countries and can be treated with antibiotics. Left untreated, however, the disease can persist for several months and sometimes becomes chronic. Chronic carriers experience no symptoms beyond their initial illness, but the bacteria remain in their cells. Controlling typhoid fever is difficult in part because carriers are unaware that they are shedding live Salmonella enterica bacteria. Some 16 million new infections occur per year.. Mark O. Clements, of the Karolinska Institute in Stockholm, Sweden, and colleagues set out to understand the genetics of typhoid fever infections. They analyzed two forms of the bacterium in mice one that causes the acute disease, and another that produces carriers. In the gene that codes for PNPase (polynucleotide phosphorylase), they discovered a single-letter mutation that shortens the protein.. Microarray analysis showed that PNPase mutants expressed more virulence factor proteins; these proteins help the ...
1E3H: A Duplicated Fold is the Structural Basis for Polynucleotide Phosphorylase Catalytic Activity, Processivity, and Regulation
Ochoa, S. and Mii, S. (1961). "Enzymatic synthesis of polynucleotides. IV. Purification and properties of polynucleotide phosphorylase from Azotobacter vinelandii". J. Biol. Chem. 236: 3303-3311. PMID 14481058. ...
Novel human polynucleotide and polypeptide sequences are disclosed that can be used in therapeutic, diagnostic, and pharmacogenomic applications.
Novel human polynucleotide and polypeptide sequences are disclosed that can be used in therapeutic, diagnostic, and pharmacogenomic applications.
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1. A new method has been developed for the preparation in good yield of highly purified Azotobacter vinelandii polynucleotide phosphorylase in its reduced form. 2. Aging or digestion with trypsin causes the enzyme to develop a primer requirement that is not eliminated by β-mercaptoethanol. 3. The development of a primer requirement is accompanied by marked changes of the electrophoretic mobility of the enzyme in polyacrylamide gels. 4. The enzyme is inactivated by aerial oxidation or thiol-specific reagents. The lost activity is restored by β-mercaptoethanol, but not by oligonucleotide primers.. ...
Involved in mRNA degradation. Catalyzes the phosphorolysis of single-stranded polyribonucleotides processively in the 3- to 5-direction.
The Gram-negative bacterium Neisseria meningitidis is a transient commensal of the human nasopharynx, but occasionally causes life-threatening disease. During colonisation of its niche, N. meningitidis has to overcome innate immune defences, including the expression of antimicrobial peptides (AMPs). Meningococcal resistance to the host defence peptide LL-37 was investigated in Papers I and II. The polysaccharide capsule and lipopolysaccharide (LPS) were found to increase LL-37 resistance by inhibiting peptide binding to the bacteria. Further, N. meningitidis responded to sub-lethal doses of LL-37 by an increase in capsule biosynthesis. Intriguingly, adhesion to epithelial cells and tissues protected N. meningitidis from physiological concentrations of LL-37 and two other helical peptides. The protective effect was mediated by RhoA- and Cdc42-dependent host cell signalling and cholesterol-rich membrane microdomains. The host epithelium thus seems to play an active role in AMP ...
A polynucleotide molecule is a biopolymer composed of 13 or more nucleotide monomers covalently bonded in a chain. DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) are examples of polynucleotides with distinct biological function. The prefix poly comes from the ancient Greek πολυς (polys, many). DNA consists of two chains of polynucleotides, with each chain in the form of a helical spiral. Although DNA and RNA do not generally occur in the same polynucleotide, the four species of nucleotides may occur in any order in the chain. The sequence of DNA or RNA species for a given polynucleotide is the main factor determining its function in a living organism or a scientific experiment. Polynucleotides occur naturally in all living organisms. The genome of an organism consists of complementary pairs of enormously long polynucleotides wound around each other in the form of a double helix. Polynucleotides have a variety of other roles in organisms. Polynucleotides are used in biochemical ...
291459338 - EP 1254270 A1 2002-11-06 - METHODS AND MATERIALS RELATING TO CUB DOMAIN POLYPEPTIDES AND POLYNUCLEOTIDES - [origin: WO0157267A1] The invention provides novel polynucleotides and polypeptides encoded by such polynucleotides and mutants or variants thereof that correspond to a novel human secreted CUB domain polypeptide. These polynucleotides comprise nucleic acid sequences isolated from cDNA library from testis (Hyseq clone identification numbers 2924342 (SEQ ID NO: 1)). Other aspects of the invention include vectors containing processes for producing novel humain secreted CUB domain polypeptides, and antibodies specific for such polypeptides.[origin: WO0157267A1] The invention provides novel polynucleotides and polypeptides encoded by such polynucleotides and mutants or variants thereof that correspond to a novel human secreted CUB domain polypeptide. These polynucleotides comprise nucleic acid sequences isolated from cDNA library from testis (Hyseq clone identification numbers 2924342 (SEQ
A composition for delivery of a polynucleotide to mucosal, neural, or other cells, comprising a GN1-binding protein and a polynucleotide in association with the binding protein; and a method for modulating immunity comprising administering the composition to an animal and expressing the polynucleotide whereby the animal generates an immune response to the product of the polynucleotide; and a method for gene therapy comprising administering to an animal a GM1-binding protein and a functional polynucleotide and expressing the polynucleotide in the animal whereby the function of the polynucleotide confers on the animal a therapeutic effect.
TY - JOUR. T1 - Helicase activity plays a crucial role for RNase R function in vivo and for RNA metabolism. AU - Tofajjen Hossain, Sk. AU - Deutscher, Murray P. PY - 2016. Y1 - 2016. N2 - RNase R is a 3 to 5 hydrolytic exoribonuclease that has the unusual ability to digest highly structured RNA. The enzyme possesses an intrinsic, ATP-dependent RNA helicase activity that is essential in vitro for efficient nuclease activity against double-stranded RNA substrates, particularly at lower temperatures, with more stable RNA duplexes, and for duplexes with short 3 overhangs. Here, we inquired whether the helicase activity was also important for RNase R function in vivo and forRNAmetabolism.Wefind that strains containing a helicasedeficient RNase R due to mutations in its ATP-binding Walker motifs exhibit growth defects at low temperatures. Most importantly, cells also lacking polynucleotide phosphorylase (PNPase), and dependent for growth on RNase R, grow extremely poorly at 34, 37, and 42 °C and ...
MAYNILA - Ikinalulungkot ng Philippine National Police Academy (PNPA) ang pagkamatay ng isa sa kanilang kadete dahil sa heatstroke.
Novel human polynucleotide and polypeptide sequences are disclosed that can be used in therapeutic, diagnostic, and pharmacogenomic applications.
TY - JOUR. T1 - Hydrolysis catalyzed with a resin containing histidine groups. AU - Hung, Wei Hsiu. AU - Hu, Cho Chun. AU - Liu, Chuen Ying. PY - 1996/1/1. Y1 - 1996/1/1. N2 - A histidine-containing polymer was synthesized in which the amino group of the histidine was attached chemically via an azide coupling method to the carboxylic acid of Amberlite IRC-50. The resultant polymer was applied as a catalyst for hydrolysis of p-nitrophenyl acetate (PNPA). PNPA in aqueous solution was hydrolyzed at 25°C with a phosphate buffer (pH 7.8). The observed kinetics obeyed Michaelis-Menten kinetics. The reaction rates at various temperature were measured. The activation parameters, preexponential factor (A) and activation energy (Ea), were 6.64 × 10-4 min-1 and 37.5 kJ mol-1 respectively. At a pH of the medium greater than 7.8, the reaction rate remained almost constant (kobs = 0.024 min-1) and seemed to be controlled by the rate of diffusion of PNPA from the bulk solution into the catalytically active ...
What is the difference between Oligonucleotide and Polynucleotide? An oligonucleotide is shorter than a polynucleotide. Oligonucleotide is composed of one or..
The invention provides methods of suppression, prevention, and/or treatment of infection by viruses. A polynucleotide comprising an immunostimulatory sequence (an ISS) is administered to an individual who is at risk of being exposed to, has been exposed to or is infected with a virus. The ISS-containing polynucleotide is administered without any antigens of the virus. Administration of the ISS-containing polynucleotide results in reduced incidence and/or severity of one or more symptoms of virus infection.
SWISS-MODEL Repository entry for C3P5L0 (PNP_BACAA), Polyribonucleotide nucleotidyltransferase. Bacillus anthracis (strain A0248)
A polynucleotide having the nucleic acid sequence according to SEQ ID NO:1, 2 or 3 or a sequence from the nucleotide numbers 167 to 1654, 1447 to 4458, 5589 to 8168, 4403 to 4984, 4924 to 5214, 5426 to 5671, 8170 to 8790, 5195 to 5409, 7730 to 7821, 5334 to 5409 or 7730 to 7821, or a continuous sequence on a polynucleotide from the nucleotide numbers 5195 to 5409 and 7730 to 7821, or 5334 to 5409 and 7730 to 7821, each referring to SEQ ID NO:1 ...
Wnt-3 polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing Wnt-3 polypeptides and polynucleotides in therapy, and diagnostic assays for such.
The invention relates to a microfabricated device and methods of using the device for analyzing and sorting polynucleotide molecules by size.
SWISS-MODEL Template Library (SMTL) entry for 6ain.2. Crystal structure of p-nitrophenol 4-monooxygenase PnpA from Pseudomonas putida DLL-E4
1512481DNAArtificial sequenceTriple mutant HIF-1a coding sequence 1atggagggcg ccggcggcgc gaacgacaag aaaaagataa gttctgaacg tcgaaaagaa 60aagtctcgag atgcagccag atctcggcga agtaaagaat ctgaagtttt ttatgagctt 120gctcatcagt tgccacttcc acataatgtg agttcgcatc ttgataaggc ctctgtgatg 180aggcttacca tcagctattt gcgtgtgagg aaacttctgg atgctggtga tttggatatt 240gaagatgaca tgaaagcaca gatgaattgc ttttatttga aagccttgga tggttttgtt 300atggttctca cagatgatgg tgacatgatt tacatttctg ataatgtgaa caaatacatg 360ggattaactc agtttgaact aactggacac agtgtgtttg attttactca tccatgtgac 420catgaggaaa tgagagaaat gcttacacac agaaatggcc ttgtgaaaaa gggtaaagaa 480caaaacacac agcgaagctt ttttctcaga atgaagtgta ccctaactag ccgaggaaga 540actatgaaca taaagtctgc aacatggaag gtattgcact gcacaggcca cattcacgta 600tatgatacca acagtaacca acctcagtgt gggtataaga aaccacctat gacctgcttg 660gtgctgattt gtgaacccat tcctcaccca tcaaatattg aaattccttt agatagcaag 720actttcctca gtcgacacag cctggatatg aaattttctt attgtgatga aagaattacc 780gaattgatgg gatatgagcc agaagaactt ttaggccgct ...
Genetic information processingTranscriptionDegradation of RNAVacB and RNase II family 3-5 exoribonucleases (TIGR00358; EC 3.1.13.1; HMM-score: 16.6) ...
0111] In a preferred embodiment, the invention provides for a method of in vitro recombination comprising: [0112] (1) obtaining modified polynucleotide fragments according any one of the method described in any one of claims 1 to 5; [0113] (2) screening some or all of said modified polynucleotides to determine which polynucleotide or polynucleotides encode a protein or proteins of interest; [0114] (3) digesting said modified polynucleotides encoding a protein or proteins of interest with restriction enzymes to form fragments with single-stranded overhangs consisting of three nucleotide residues or of nucleotide residues in multiples of three; [0115] (4) modifying the obtained polynucleotide fragments of step (3) by removing and/or filling in the single-stranded overhangs to obtain new modified fragments by [0116] (i) removing, in multiples of three, all of the nucleotide residues of said overhanging end of one or more polynucleotide fragments; or [0117] (ii) extending the single strand of the ...
Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction. The devices comprise a substrate microfabricated to define a sample inlet port and a mesoscale flow system, which extends from the inlet port. The mesoscale flow system includes a polynucleotide polymerization reaction chamber in fluid communication with the inlet port which is provided with reagents required for polymerization and amplification of a preselected polynucleotide. In one embodiment the devices may be utilized to implement a polymerase chain reaction (PCR) in the reaction chamber (PCR chamber). The PCR chamber is provided with the sample polynucleotide, polymerase, nucleoside triphosphates, primers and other reagents required for the polymerase chain reaction, and the device is provided with a device for thermally controlling the temperature of the contents of the reaction chamber at a temperature controlled to dehybridize double stranded polynucleotide, to
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The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.
Compositions having polynucleotides encoding multiple translational stop signals in more than one reading frame are provided. The compositions include isolated polynucleotides, expression cassettes, a
The invention relates to a microfabricated device and methods of using the device for analyzing and sorting polynucleotide molecules by size.
I have seen marks denoting the number of nucleotides along the gel which actually denote the molecular weight. But , do the polynucleotides with same number of nucleotides have same molecular wt.always? I think , they would not , as the proportions of A ,T ,G & C would not be the same always.Thus , they will get separated during electrophoresis . So, how can we have a common mark denoting the nucleotide number for such polynucleotides as they would be apart from each other . Can we have only sizewise separation of molecules in electrophoresis? If yes , then ,I think , such polynucleotides may have same mark as the size-difference between them seems to be lesser ...
Zhuk R.A.; Berzinya A.E.; Silinya V.N.; Liepinsh E.E.; Giller S.A. Analogs of pyrimidine mono- and polynucleotides - 7. 2-(1-Uracilyl)tetrahydrofuran-5-carboxylic acids and their derivatives. Chem. Heterocycl. Compd. 1979, 15(8), 926-929 ...
The cold acclimatization response in many bacterial species is a tightly regulated process, which ensures the correct folding of macromolecules. In enterobacteria, this response is in part dependent on polynucleotide phosphorylase, which is encoded by the gene pnp. Based on transcriptional analysis of the pnp locus of Salmonella enterica serovar Typhimurium, we show that pnp and the adjacent membrane lipoprotein nlpI gene form an operon with both genes contributing independently to the cold acclimatization response at 15 °C. Our findings thereby define a new role for NlpI in bacterial cold acclimatization.. ...
Nucleic acid therapeutics involves the use of polynucleotides (DNA, RNA) as novel therapeutic agents for the treatment of a wide range of diseases including cancer and several metabolic and genetic disorders. However, the highly unstable nature of RNA molecules necessitates the use of drug carriers to prevent them from nuclease degradation and facilitate targeted delivery in vivo. Hence, this study was conducted to optimize the preparation of nanoparticle carriers in order to improve the stability of the polynucleotides (siRNA and mRNA). Additionally, as heterogeneity and stability of nanoparticle formulations are major issues preventing the clinical approval of therapeutic formulations this study was also focused on improving the homogeneity and the stability of the nanoparticles. In the siRNA study, reconstituted high density lipoprotein (rHDL) nanoparticles were used as the delivery vector. Optimization of siRNA-rHDL formulation was attempted with respect to homogeneity, size of the nanoparticle and
Globally modulates RNA abundance by binding to RNase E (Rne) and regulating its endonucleolytic activity. Can modulate Rne action in a substrate-dependent manner by altering the composition of the degradosome. Modulates RNA-binding and helicase activities of the degradosome.
casSAR Dugability of Q89WK4 | rph | Ribonuclease PH - Also known as RNPH_BRADU, rph. Phosphorolytic 3-5 exoribonuclease that plays an important role in tRNA 3-end maturation. Removes nucleotide residues following the 3-CCA terminus of tRNAs; can also add nucleotides to the ends of RNA molecules by using nucleoside diphosphates as substrates, but this may not be physiologically important. Probably plays a role in initiation of 16S rRNA degradation (leading to ribosome degradation) during starvation. Homohexameric ring arranged as a trimer of dimers.
casSAR Dugability of C3K479 | rph | Ribonuclease PH - Also known as RNPH_PSEFS, rph. Phosphorolytic 3-5 exoribonuclease that plays an important role in tRNA 3-end maturation. Removes nucleotide residues following the 3-CCA terminus of tRNAs; can also add nucleotides to the ends of RNA molecules by using nucleoside diphosphates as substrates, but this may not be physiologically important. Probably plays a role in initiation of 16S rRNA degradation (leading to ribosome degradation) during starvation. Homohexameric ring arranged as a trimer of dimers.
The invention relates to methods for pairwise sequencing of a double-stranded polynucleotide template, which permit the sequential determination of nucleotide sequences in two distinct and separate r
Verify DIPHOSPHATES in Scrabble dictionary and games, check DIPHOSPHATES definition, DIPHOSPHATES in wwf, Words With Friends score for DIPHOSPHATES, definition of DIPHOSPHATES.
In early physical studies of DNA, a variety of experiments indicated that DNA molecules occur in long helixes with each helix being formed from two or more polynucleotide chains bound side by side. Chemical analyses also demonstrated that the phosphate groups were on the outside of the helix and that the number of A and T residues in DNA were always equal, as were those of G and C. With these facts in mind, Watson and Crick in 1953 proposed that the DNA molecule actually consists of two polynucleotide chains coiled around the same axis to form a double helix (Figure 3). In this model, the hydrophilic sugar-phosphate groups follow the outer edges of the molecule where they can interact with water. The hydrophobic bases face inward toward each other in the molecules center and thus avoid contact with water. The two polynucleotide strands run in opposite directions (they are anti- parallel) and are held together primarily by hydrogen and hydrophobic bonding between the bases, where A is always ...