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1.
Radiat Prot Dosimetry ; 146(1-3): 198-201, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21498409

ABSTRACT

To predict how accurately neutron dosemeters can measure the neutron dose equivalent (rate) in MOX fuel fabrication facility work environments, the dose equivalent responses of neutron dosemeters were calculated by the spectral folding method. The dosemeters selected included two types of personal dosemeter, namely a thermoluminescent albedo neutron dosemeter and an electronic neutron dosemeter, three moderator-based neutron survey meters, and one special instrument called an H(p)(10) monitor. The calculations revealed the energy dependences of the responses expected within the entire range of neutron spectral variations observed in neutron fields at workplaces.


Subject(s)
Neutrons , Occupational Exposure , Equipment Design , Humans , Radiation Dosage , Radiation Monitoring/instrumentation , Radiation Protection/instrumentation , Thermoluminescent Dosimetry/instrumentation
2.
Radiat Prot Dosimetry ; 144(1-4): 575-9, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21062798

ABSTRACT

At the Nuclear Fuel Cycle Engineering Laboratories of the Japan Atomic Energy Agency (JAEA), small pieces of indium foil incorporated into personal dosemeters have been used for personnel screening in criticality accidents. Irradiation tests of the badges were performed using the SILENE reactor to verify the calibration of the indium activation that had been made in the 1980s and to recalibrate them for simulated criticalities that would be the most likely to occur in the solution process line. In addition, Monte Carlo calculations of the indium activation using the badge model were also made to complement the spectral dependence. The results lead to a screening level of 15 kcpm being determined that corresponds to a total dose of 0.25 Gy, which is also applicable in posterior-anterior exposure. The recalibration based on the latest study will provide a sounder basis for the screening procedure in the event of a criticality accident.


Subject(s)
Indium/analysis , Radiation Monitoring/instrumentation , Radiation Protection/instrumentation , Radioactive Hazard Release , Radiometry/instrumentation , Calibration , Computer Simulation , Humans , Monte Carlo Method , Neutrons , Radiation Dosage , Radiation Monitoring/methods , Radiation Protection/methods , Radiometry/methods , Silene/chemistry , Thermoluminescent Dosimetry/instrumentation , Thermoluminescent Dosimetry/methods
3.
Radiat Prot Dosimetry ; 126(1-4): 261-4, 2007.
Article in English | MEDLINE | ID: mdl-17545659

ABSTRACT

A new neutron-measuring instrument that is intended to measure a neutron personal dose equivalent, H(p)(10) was developed. This instrument is composed of two parts: (1) a conventional moderator-based neutron dose equivalent meter and (2) a neutron shield made of borated polyethylene, which covers a backward hemisphere to adjust the angular dependence. The whole design was determined on the basis of MCNP calculations so as to have response characteristics that would generally match both the energy and angular dependencies of H(p)(10). This new instrument will be a great help in assessing the reference values of neutron H(p)(10) during field testing of personal neutron dosemeters in workplaces and also in interpreting their readings.


Subject(s)
Computer-Aided Design , Neutrons , Radiation Monitoring/instrumentation , Radiation Protection/instrumentation , Equipment Design , Equipment Failure Analysis , Humans , Monte Carlo Method , Radiation Dosage , Radiation Monitoring/methods , Radiation Protection/methods , Reproducibility of Results , Sensitivity and Specificity
4.
Radiat Prot Dosimetry ; 127(1-4): 411-4, 2007.
Article in English | MEDLINE | ID: mdl-17567761

ABSTRACT

The aim of this study is to propose action levels for chelation therapy in the case of inhalation of plutonium compounds using nose swabs. The relationship between the activity found in the nose swabs and early faecal excretion was investigated using actual cases at JAEA-NFCEL. The ratio was found to be in log-normal distribution. The action levels based on the activity of nose swab corresponding to 10 ALI (=200 mSv) are determined for the facilities at JAEA-NFCEL by using the relationship and specific information such as isotopic ratio and physicochemical characteristics of plutonium compounds.


Subject(s)
Biological Assay/methods , Chelating Agents/therapeutic use , Nasal Mucosa/metabolism , Plutonium/administration & dosage , Plutonium/pharmacokinetics , Radiation Injuries/prevention & control , Radiometry/methods , Body Burden , Computer Simulation , Humans , Maximum Allowable Concentration , Models, Biological , Plutonium/toxicity , Radiation Dosage , Radiation Injuries/etiology , Relative Biological Effectiveness , Reproducibility of Results , Sensitivity and Specificity
5.
Radiat Prot Dosimetry ; 125(1-4): 383-6, 2007.
Article in English | MEDLINE | ID: mdl-17337735

ABSTRACT

The authors carried out an operational study that compared the use of TLD albedo dosemeters and solid state nuclear tracks detector in plutonium environments of Japan Nuclear Cycle Development Institute, Tokai Works. A selected group of workers engaged in the fabrication process of MOX (Plutonium-Uranium mixed oxide) fuel wore both TLD albedo dosemeters and solid state nuclear tracks detectors. The TL readings were generally proportional to the counted etch-pits, and thus the dose equivalent results obtained from TLD albedo dosemeter agreed with those from solid state nuclear tracks detector within a factor of 1.5. This result indicates that, in the workplaces of the MOX fuel plants, the neutron spectrum remained almost constant in terms of time and space, and the appropriate range of field-specific correction with spectrum variations was small in albedo dosimetry.


Subject(s)
Occupational Exposure/analysis , Power Plants , Radiation Monitoring/instrumentation , Radiation Protection/instrumentation , Equipment Design , Equipment Failure Analysis , Internationality , Japan , Linear Energy Transfer , Radiation Dosage , Reproducibility of Results , Semiconductors , Sensitivity and Specificity
6.
Br J Dermatol ; 151(6): 1133-42, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15606507

ABSTRACT

BACKGROUND: Olopatadine hydrochloride (olopatadine) is one of the second-generation antihistamines, which is prescribed for allergic disorders such as rhinitis, urticaria and eczema dermatitis. OBJECTIVES: To investigate the possible anti-inflammatory effect of olopatadine on the chronic contact hypersensitivity response to repeated topical application of oxazolone in mice. METHODS: The preventive and therapeutic effects of oral olopatadine were quantified by measurements of ear swelling, cytokine protein and mRNA expression in the ear lesion, and were compared with those of topical betamethasone 17-valerate (betamethasone). RESULTS: The ear receiving repeated applications of oxazolone exhibited erythema, oedema and abrasion. Both preventive and therapeutic administration of olopatadine (10 mg kg(-1) day(-1)) significantly inhibited the ear swelling and the increased production of interleukin (IL)-4, IL-1beta, granulocyte-macrophage colony-stimulating factor (GM-CSF) and nerve growth factor. In the histopathological analysis, olopatadine ameliorated epidermal hyperplasia and infiltration of inflammatory cells. Consistent with these results, olopatadine significantly reduced the increased expression of interferon-gamma and IL-4 mRNA. Although betamethasone (0.012 mg ear(-1) day(-1)) showed similar activities to olopatadine against these responses, it caused atrophy of the ear skin. CONCLUSIONS: These results indicate that olopatadine is an antihistamine agent having inhibitory activities against chronic inflammatory dermatitis, possibly resulting from its diminishing effect on elevated cytokines.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Dermatitis, Contact/drug therapy , Dibenzoxepins/therapeutic use , Histamine H1 Antagonists, Non-Sedating/therapeutic use , Adjuvants, Immunologic , Animals , Chronic Disease , Cytokines/biosynthesis , Cytokines/genetics , Dermatitis, Contact/etiology , Dermatitis, Contact/immunology , Dermatitis, Contact/prevention & control , Gene Expression Regulation/drug effects , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Immunoglobulin E/blood , Male , Mice , Mice, Inbred BALB C , Nerve Growth Factor/biosynthesis , Olopatadine Hydrochloride , Oxazolone , RNA, Messenger/genetics
7.
Tech Coloproctol ; 6(2): 121-3, 2002 Sep.
Article in English | MEDLINE | ID: mdl-12402059

ABSTRACT

We report pneumatosis cystoides intestinalis in a 10-month-old girl who developed bloody diarrhea following chemotherapy for leukemia. The diagnosis was made only by colonic endoscopic ultrasonography, whereas the abdominal plain radiogram and computed tomography failed to elucidate the diagnosis. She was successfully treated with hyperbaric oxygen therapy. Wider application of endoscopic ultrasonography may lead to the more frequent detection of pneumatosis cystoides intestinalis, currently a rare disorder.


Subject(s)
Endosonography , Pneumatosis Cystoides Intestinalis/diagnostic imaging , Female , Humans , Hyperbaric Oxygenation , Infant , Pneumatosis Cystoides Intestinalis/therapy , Radiography
8.
Biosci Biotechnol Biochem ; 65(8): 1824-31, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11577723

ABSTRACT

The intracellular beta-xylosidase was induced when Streptomyces thermoviolaceus OPC-520 was grown at 50 degrees C in a minimal medium containing xylan or xylooligosaccharides. The 82-kDa protein with beta-xylosidase activity was partially purified and its N-terminal amino acid sequence was analyzed. The gene encoding the enzyme was cloned, sequenced, and expressed in Escherichia coli. The bxlA gene consists of a 2,100-bp open reading frame encoding 770 amino acids. The deduced amino acid sequence of the bxlA gene product had significant similarity with beta-xylosidases classified into family 3 of glycosyl hydrolases. The bxlA gene was expressed in E. coli, and the recombinant protein was purified to homogeneity. The enzyme was a monomer with a molecular mass of 82 kDa. The purified enzyme showed hydrolytic activity towards only p-nitrophenyl-beta-D-xylopyranoside among the synthetic glycosides tested. Thin-layer chromatography analysis showed that the enzyme is an exo-type enzyme that hydrolyze xylooligosaccharides, but had no activity toward xylan. High activity against pNPX occurred in the pH range 6.0-7.0 and temperature range 40-50 degrees C.


Subject(s)
Streptomyces/enzymology , Streptomyces/genetics , Xylosidases/genetics , Amino Acid Sequence , Base Sequence , Chromosomes, Bacterial/enzymology , Cloning, Molecular , Culture Media , Electrophoresis, Polyacrylamide Gel , Escherichia coli/enzymology , Genes, Bacterial , Hydrolysis , Molecular Sequence Data , Plasmids/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/isolation & purification , Xylosidases/biosynthesis
9.
RNA ; 7(8): 1153-64, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11497433

ABSTRACT

Rpp21, a protein subunit of human nuclear ribonuclease P (RNase P) was cloned by virtue of its homology with Rpr2p, an essential subunit of Saccharomyces cerevisiae nuclear RNase P. Rpp21 is encoded by a gene that resides in the class I gene cluster of the major histocompatibility complex, is associated with highly purified RNase P, and binds precursor tRNA. Rpp21 is predominantly localized in the nucleoplasm but is also observed in nucleoli and Cajal bodies when expressed at high levels. Intron retention and splice-site selection in Rpp21 precursor mRNA regulate the intranuclear distribution of the protein products and their association with the RNase P holoenzyme. Our study reveals that dynamic nuclear structures that include nucleoli, the perinucleolar compartment and Cajal bodies are all involved in the production and assembly of human RNase P.


Subject(s)
Endoribonucleases/chemistry , RNA, Catalytic/chemistry , 3T3 Cells , Alternative Splicing , Amino Acid Sequence , Animals , Blotting, Western , Cell Nucleus/chemistry , Cell Nucleus/metabolism , Cells, Cultured , Cloning, Molecular , DNA, Complementary/metabolism , Endoribonucleases/metabolism , Fibroblasts/metabolism , HeLa Cells , Humans , Introns , Major Histocompatibility Complex , Mice , Microscopy, Fluorescence , Models, Genetic , Molecular Sequence Data , Precipitin Tests , Protein Binding , RNA Splicing , RNA, Catalytic/metabolism , RNA, Messenger/metabolism , RNA, Transfer/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Ribonuclease P , Saccharomyces cerevisiae/chemistry , Saccharomyces cerevisiae/genetics , Sequence Homology, Amino Acid
10.
RNA ; 7(7): 937-41, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11455963

ABSTRACT

A yeast three-hybrid system was employed to analyze interactions in vivo between H1 RNA, the RNA subunit of human nuclear RNase P, and eight of the protein subunits of the enzyme. The genetic analysis indicates that subunits Rpp21, Rpp29, Rpp30, and Rpp38 interact directly with H1 RNA. The results of direct UV crosslinking studies of the purified RNase P holoenzyme confirm the results of the three-hybrid assay.


Subject(s)
Cell Nucleus/enzymology , Endoribonucleases/metabolism , RNA, Catalytic/metabolism , RNA-Binding Proteins/metabolism , RNA/metabolism , Blotting, Western , Endoribonucleases/chemistry , HeLa Cells , Humans , RNA, Catalytic/chemistry , Ribonuclease P
11.
Proc Natl Acad Sci U S A ; 98(12): 6605-10, 2001 Jun 05.
Article in English | MEDLINE | ID: mdl-11381134

ABSTRACT

Narrow spectrum antimicrobial activity has been designed to reduce the expression of two essential genes, one coding for the protein subunit of RNase P (C5 protein) and one for gyrase (gyrase A). In both cases, external guide sequences (EGS) have been designed to complex with either mRNA. Using the EGS technology, the level of microbial viability is reduced to less than 10% of the wild-type strain. The EGSs are additive when used together and depend on the number of nucleotides paired when attacking gyrase A mRNA. In the case of gyrase A, three nucleotides unpaired out of a 15-mer EGS still favor complete inhibition by the EGS but five unpaired nucleotides do not.


Subject(s)
Bacterial Proteins/genetics , DNA Topoisomerases, Type II/genetics , Endoribonucleases/genetics , Escherichia coli Proteins , Escherichia coli/genetics , Genes, Bacterial/physiology , RNA, Catalytic/genetics , Base Sequence , DNA Gyrase , Escherichia coli/physiology , Molecular Sequence Data , RNA, Messenger/metabolism , RNA, Transfer/metabolism , Ribonuclease P
13.
Mutat Res ; 444(1): 133-44, 1999 Jul 21.
Article in English | MEDLINE | ID: mdl-10477347

ABSTRACT

Two classes of the free radical Maillard intermediates, the pyrazine cation radical and the carbon-centered radicals, are detected in the reaction of Glc (glucose)/Gly (glycine) by electron spin resonance and spin trapping technique. Profile of the generation of the pyrazine cation radical in the reaction with different ratios of the reactants was found to be similar to that of the formation of mutagens in the subsequent reaction with creatinine. By contrast, profile of the generation of the carbon-centered radicals was not consistent with that of the mutagen formation. Thiol antioxidants and unsaturated fatty acids (or their esters) effectively scavenged the pyrazine cation radical generated in the reaction of Glc/Gly, and inhibited the formation of the mutagens in the reaction of Glc/Gly and creatinine. Ethanol, a sulfide and a saturated fatty acid were not effective to scavenge the pyrazine cation radical and did not inhibit the mutagen formation. The pyrazine cation radical rather than the carbon-centered radicals may play an important role in the mutagen formation. Thiol antioxidants and unsaturated fatty acids can be evaluated as inhibitors of the pyrazine cation radical-derived formation of the mutagens.


Subject(s)
Creatinine/metabolism , Glucose/metabolism , Glycine/metabolism , Mutagens/metabolism , Cations , Electron Spin Resonance Spectroscopy , Food Analysis , Free Radicals , Hot Temperature , Maillard Reaction , Mutagenicity Tests , Mutagens/chemistry , Mutagens/toxicity , Pyrazines/chemistry , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics
14.
RNA ; 4(4): 407-17, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9630247

ABSTRACT

At least six proteins co-purify with human ribonuclease P (RNase P), a tRNA processing ribonucleoprotein. Two of these proteins, Rpp30 and Rpp38, are Th autoantigens. Recombinant Rpp30 and Rpp38 are also recognized by Th sera from systemic sclerosis patients. Two of the other proteins associated with RNase P, Rpp20 and Rpp40, do not cross-react with Th sera. Polyclonal antibodies raised against all four recombinant proteins recognize the corresponding proteins associated with RNase P and precipitate active holoenzyme. Catalytically active RNase P holoenzyme can be separated from the nucleolar and mitochondrial RNA processing endoribonuclease, RNase MRP, even though these two enzymes may share some subunits.


Subject(s)
Autoantigens/immunology , Carrier Proteins , Endoribonucleases/immunology , RNA, Catalytic/immunology , Ribonucleoproteins/immunology , Amino Acid Sequence , Apoptosis Regulatory Proteins , Autoantigens/biosynthesis , Autoantigens/genetics , Base Sequence , Cloning, Molecular , Conserved Sequence , Endoribonucleases/biosynthesis , Endoribonucleases/genetics , Eukaryotic Cells , Humans , Molecular Sequence Data , Precipitin Tests , RNA, Catalytic/biosynthesis , RNA, Catalytic/genetics , Recombinant Proteins/biosynthesis , Recombinant Proteins/immunology , Ribonuclease P , Ribonucleoproteins/biosynthesis , Ribonucleoproteins/genetics
15.
Proc Natl Acad Sci U S A ; 94(16): 8468-72, 1997 Aug 05.
Article in English | MEDLINE | ID: mdl-9238000

ABSTRACT

Plasmids that contain synthetic genes coding for small oligoribonucleotides called external guide sequences (EGSs) have been introduced into strains of Escherichia coli harboring antibiotic resistance genes. The EGSs direct RNase P to cleave the mRNAs transcribed from these genes thereby converting the phenotype of drug-resistant cells to drug sensitivity. Increasing the EGS-to-target mRNA ratio by changing gene copy number or the number of EGSs complementary to different target sites enhances the efficiency of the conversion process. We demonstrate a general method for the efficient phenotypic conversion of drug-resistant bacterial cultures.


Subject(s)
Drug Resistance/genetics , Endoribonucleases/genetics , Escherichia coli Proteins , Escherichia coli/genetics , Gene Expression Regulation, Bacterial , Phenotype , RNA, Catalytic/genetics , Plasmids/genetics , RNA, Messenger/genetics , Ribonuclease P , Transfection
16.
Radiat Med ; 14(3): 151-4, 1996.
Article in English | MEDLINE | ID: mdl-8827810

ABSTRACT

A retrospective analysis was performed to investigate the radioprotective effects of azelastine against radiation dermatitis for patients with head and neck cancers. The effects of azelastine were studied in 19 patients with laryngeal cancers treated by irradiation. As controls, 29 patients with laryngeal cancers treated by irradiation without the administration of azelastine were studied. All patients were irradiated using 3 MV linac X-rays. Azelastine was administered orally twice a day. Moist desquamation was observed in four of 29 control patients whereas no such moist desquamation developed after the administration of azelastine. Two cases of moist desquamation that developed before the administration of azelastine regressed during irradiation in patients placed on azelastine. Radiotherapy was completed without interruption in all patients treated with azelastine. No severe side effects were observed. Azelastine, administered orally, was a safe drug and has the potential of improving skin tolerance in irradiation therapy.


Subject(s)
Phthalazines/therapeutic use , Radiation-Protective Agents/therapeutic use , Radiodermatitis/drug therapy , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Head and Neck Neoplasms/radiotherapy , Humans , Retrospective Studies , Urticaria/prevention & control
17.
Radiat Med ; 13(6): 297-300, 1995.
Article in English | MEDLINE | ID: mdl-8850371

ABSTRACT

From 1980 to 1990, a total of 54 patients with prostatic carcinoma were treated with external radiation therapy at the Kumamoto National Hospital. Ten patients were classified as Stage B, 22 as Stage C, and another 22 as Stage D according to the American Urological Association Clinical Staging System. The 5-year survival for all 54 patients was 30%. The 5-year disease-specific survival was 67% for Stage B, 47% for Stage C, and 26% for Stage D. The 5-year survival was 43% for patients in whom radiation therapy was initiated immediately after the first diagnosis or with less than one year of hormonal therapy, while it was 0% for patients in whom radiation therapy was initiated after more than one year of hormonal therapy (p = 0.01). The cause of intercurrent death was acute myocardial infarction in four patients and acute cardiac failure in one. Four of these patients received hormonal therapy for more than one year. The incidence of radiation-induced proctitis was not severe. This study suggests that long-term hormonal therapy prior to radiation therapy worsens the prognosis of patients with prostatic carcinoma.


Subject(s)
Antineoplastic Agents, Hormonal/therapeutic use , Carcinoma/radiotherapy , Prostatic Neoplasms/radiotherapy , Aged , Aged, 80 and over , Antineoplastic Agents/therapeutic use , Carcinoma/drug therapy , Cardiac Output, Low/etiology , Cause of Death , Combined Modality Therapy , Diethylstilbestrol/analogs & derivatives , Diethylstilbestrol/therapeutic use , Disease-Free Survival , Estramustine/therapeutic use , Follow-Up Studies , Humans , Male , Middle Aged , Myocardial Infarction/etiology , Neoplasm Staging , Proctitis/etiology , Prognosis , Prostatic Neoplasms/drug therapy , Radiation Injuries/etiology , Radiotherapy Dosage , Retrospective Studies , Survival Rate
18.
Proc Natl Acad Sci U S A ; 92(24): 11115-9, 1995 Nov 21.
Article in English | MEDLINE | ID: mdl-7479948

ABSTRACT

Plasmids encoding various external guide sequences (EGSs) were constructed and inserted into Escherichia coli. In strains harboring the appropriate plasmids, the expression of fully induced beta-galactosidase and alkaline phosphatase activity was reduced by more than 50%, while no reduction in such activity was observed in strains with non-specific EGSs. The inhibition of gene expression was virtually abolished at restrictive temperatures in strains that were temperature-sensitive for RNase P (EC 3.1.26.5). Northern blot analysis showed that the steady-state copy number of EGS RNAs was several hundred per cell in vivo. A plasmid that contained a gene for M1 RNA covalently linked to a specific EGS reduced the level of expression of a suppressor tRNA that was encoded by a separate plasmid. Similar methods can be used to regulate gene expression in E. coli and to mimic the properties of cold-sensitive mutants.


Subject(s)
Endoribonucleases/physiology , Escherichia coli Proteins , Escherichia coli/genetics , Gene Expression Regulation, Bacterial , RNA, Catalytic/physiology , Base Sequence , Escherichia coli/enzymology , Molecular Sequence Data , Oligoribonucleotides/chemistry , RNA, Antisense/chemistry , RNA, Bacterial/genetics , RNA, Messenger/genetics , Ribonuclease P , Temperature
19.
Kansenshogaku Zasshi ; 68(9): 1113-6, 1994 Sep.
Article in Japanese | MEDLINE | ID: mdl-7963802

ABSTRACT

A case of streptococcal toxic shock-like syndrome in a previously healthy, 57 year old Japanese female has been reported. Initially, she had a sore throat and low grade fever for 5 days. Because of sudden severe pain on the extremities and erythema on bilateral forearms, she was hospitalized. On admission, her conciousness was clear. Although profound hypotension, anuria and prolonged blood coagulation were observed. Antibiotics, fluid therapy and dopamine were given. Four hours after admission, she died in spite of resuscitation efforts, by sudden cardiac arrest. Streptococcus pyogenes was isolated in her blood. At the same time as when she died, three of the five people of the patient's family living with her, had pharingitis or pneumonia. From the pharynxs of the three people with pharingitis, Streptococcus pyogenes was also isolated. The serotype of all organisms was T11, and they produced exotoxintype B in vitro. This case suggests that infection of Streptococcus pyogenes is not essential for the development of toxic shock-like syndrome.


Subject(s)
Pharyngitis/etiology , Shock, Septic/complications , Streptococcal Infections/etiology , Streptococcus pyogenes , Family Health , Fatal Outcome , Female , Humans , Middle Aged
20.
Biochemistry ; 32(28): 7152-61, 1993 Jul 20.
Article in English | MEDLINE | ID: mdl-7688247

ABSTRACT

A gel-shift assay was devised to detect stable enzyme-substrate (E-S) complexes between M1 RNA, the catalytic subunit of RNase P from Escherichia coli, and its tRNA precursor substrates. The use of deletion derivatives of M1 RNA in the gel-shift assay has allowed us to identify regions of the enzyme that are involved in the binding of the substrate or that are necessary for catalytic activity. Fragments of substrates that contain the 3' CCA sequence bind preferentially to regions in the 5' half of M1 RNA, while 5' leader sequences interact primarily with regions in the 3' half of M1 RNA. The 5' leader sequence present in the precursor to tRNA(Tyr)su3 from E. coli plays an important role in the formation of stable E-S complexes with M1 RNA. The CCA sequence at the 3' end of precursor tRNA substrates is involved in the product-release step of the reaction that is catalyzed by M1 RNA. Direct measurements of the concentrations of all the components in the reaction catalyzed by M1 RNA facilitated a new approach to the kinetic analysis of the action of the enzyme.


Subject(s)
Endoribonucleases/metabolism , Escherichia coli Proteins , RNA Precursors/metabolism , RNA, Bacterial/metabolism , RNA, Catalytic/metabolism , RNA, Transfer/metabolism , Base Sequence , Electrophoresis, Agar Gel , Escherichia coli/metabolism , Kinetics , Metals/pharmacology , Molecular Sequence Data , Nucleic Acid Conformation , RNA, Bacterial/drug effects , Ribonuclease P , Substrate Specificity
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