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1.
Differential Thermolability of Exonuclease and Endonuclease Activities of the recBC Nuclease Isolated from Thermosensitive recB and recC Mutants 总被引:15,自引:10,他引:5 下载免费PDF全文
Sidney R. Kushner 《Journal of bacteriology》1974,120(3):1219-1222
The recBC nuclease (also called exonuclease V) has been partially purified from Escherichia coli K-12 strains carrying the thermosensitive recB270, recC271, and recB270 recC271 mutations. Of the multiple activities associated with the enzyme, only the adenosine 5'-triphosphate-dependent exonucleolytic hydrolysis of duplex deoxyribonucleic acid (DNA) is abnormally thermolabile. The exo- and endonucleolytic degradation of single-stranded DNA is no more thermosensitive than that catalyzed by the wild-type enzyme. These results suggest that the defects in genetic recombination, DNA repair, and the maintenance of cell viability observed in recBC mutants in vivo result primarily from the specific loss of adenosine 5'-triphosphate-dependent exonuclease active on duplex DNA. 相似文献
2.
Calcium and the mechanism of axoplasmic transport 总被引:2,自引:0,他引:2
S Ochs 《Federation proceedings》1982,41(7):2301-2306
Using desheathed cat peroneal nerves in in vitro studies, Ca2+ was recently shown to be required to maintain axoplasmic transport. Calmodulin was also shown to be present in nerve and to participate in transport. These findings open up new possibilities for a better understanding of the underlying mechanism of transport. In the transport filament model, the materials transported are bound to a common carrier, the transport filaments, which are moved along the microtubules by means of an interaction with the side arms of the microtubules. This is an energy-requiring process that depends on a supply of ATP, which is utilized by the Ca2+,Mg2+-ATPase associated with the side arms of the microtubules. The Ca2+,Mg2+-ATPase is activated by calmodulin at the low micromolar levels of free Ca2+ present in the axon. The level is kept low by calcium-regulatory mechanisms that include mitochondria, endoplasmic reticulum, and calcium-binding proteins. Nerves exposed to higher-than-normal concentrations of Ca2+ in the medium show an increased number of particles in these organelles as expected of their Ca2+-regulatory role. The nature of the calmodulin-Ca,Mg-ATPase complex associated with the side arms is discussed on the basis of the transport model. Also discussed is slow transport, which is explained on the basis of the model as a differential binding affinity to the transport filaments. 相似文献
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The three potent antimitotic vinca alkaloids: vincristine (VCR), vinblastine (VLB), and vindesine (VDS) were compared for their effect in blocking axoplasmic transport in vitro using a desheathed preparation of the peroneal branch of cat sciatic nerve. A range of vinca alkaloid concentrations from 1–100μM was examined. The relative order of potency in blocking axoplasmic transport was VCR > VLB > VDS at a concentration of 25μM. At the higher concentrations block occurred so rapidly that a statistically significant difference between these agents could not be obtained. The relation of vinca block ot the transport mechanism is discussed. 相似文献
7.
Retroperitoneal fibromatosis and acquired immunodeficiency syndrome in macaques: clinical and immunologic studies 总被引:5,自引:0,他引:5
C C Tsai W E Giddens H D Ochs W R Morton G H Knitter G A Blakley R E Benveniste 《Laboratory animal science》1986,36(2):119-125
A simian acquired immunodeficiency syndrome (SAIDS) associated with retroperitoneal fibromatosis (RF) has been observed in several species of macaque at the Washington Regional Primate Research Center. Clinical signs were recurrent diarrhea, weight loss, mesenteric lymphadenopathy, and opportunistic infections. Most affected macaques in the later stages of illness showed marked immunodeficiency. Response of peripheral blood mononuclear cells to mitogens was impaired significantly. There was sharply depressed primary and secondary antibody response to the T-cell dependent antigen, bacteriophage phi X174. Affected monkeys did not switch from IgM to IgG antibody following a secondary immunization, as did normal macaques. Twenty-four (67%) of 36 affected animals with progressive RF or deteriorated stages of illness had hypoproteinemia and hypoalbuminemia. Quantitative serum immunoglobulins of 23 cases showed that eight (35%) had hypogammaglobulinemia, six (26%) had hypergammaglobulinemia, and the remainder (39%) were within the normal range. Opportunistic infections were predominantly bacterial pathogens. Type D retrovirus appeared to be closely associated with RF-affected macaques (12/12 or 100%). The case fatality rate (including animals sacrificed after prolonged illness) was 98%. The leading cause of death was due directly to RF lesions in 43%, to enterocolitis in 36%, septicemia in 12%, amyloidosis in 5%, and malignant lymphoma (2%). Clinical, immunologic and pathologic changes reveal an acquired immunodeficiency syndrome that has many similarities to human AIDS. SAIDS and RF may be a useful model for studying human AIDS. 相似文献
8.
Glutamine metabolism of isolated rat hepatocytes. Evidence for catecholamine activation of alpha-ketoglutarate dehydrogenase 总被引:2,自引:0,他引:2
R S Ochs 《The Journal of biological chemistry》1984,259(21):13004-13010
Effects of norepinephrine on gluconeogenesis and ureogenesis from glutamine by hepatocytes from fasted rats were assessed. Comparisons were made to asparagine metabolism and to the effects of NH4Cl and dibutyryl cyclic AMP. With asparagine as substrate, aspartate content was very high but norepinephrine, dibutyryl cyclic AMP, or NH4Cl had little effect on gluconeogenesis or ureogenesis. Metabolism of asparagine could be greatly enhanced by the combination of oleate, ornithine, and NH4Cl. However, even under these conditions, asparatate content remained high, and norepinephrine and dibutyryl cyclic AMP had little influence on glucose or urea synthesis. With glutamine as substrate, aspartate content was much lower, but was greatly elevated by norepinephrine, dibutyryl cyclic AMP, or NH4Cl. Each of these effectors strongly stimulated glucose and urea formation from glutamine. NH4Cl stimulation was accompanied by an increased glutamate and decreased alpha-ketoglutarate content. This suggests the mechanism for NH4Cl stimulation is a near-equilibrium adjustment to ammonia by glutamate dehydrogenase and aspartate aminotransferase rather than a principal involvement of glutaminase. Although both norepinephrine and dibutyryl cyclic AMP lowered alpha-ketoglutarate to the same extent, norepinephrine more rapidly increased aspartate content and led to a smaller accumulation of glutamate than did dibutyryl cyclic AMP. Moreover, only norepinephrine led to a rapid increase in succinyl-CoA concentration. The catecholamine effect could not be explained by specific changes in cytosolic or mitochondrial redox states. The results suggest that alpha-ketoglutarate dehydrogenase is a site of catecholamine action in rat liver. Since purified alpha-ketoglutarate dehydrogenase is known to be Ca2+ stimulated and Ca2+ flux is involved in catecholamine action, these findings also suggest that mitochondrial Ca2+ is elevated by catecholamines. 相似文献
9.
In vivo chromosomal instability in ataxia-telangiectasia homozygotes and heterozygotes 总被引:1,自引:0,他引:1
Summary The exfoliated cell micronucleus test was used to monitor in vivo chromosomal instability in a population comprised of five ataxia-telangiectasia (A-T) homozygotes and seven obligate heterozygotes (parents of A-T patients). This assay was previously validated as a procedure for quantifying non-invasively carcinogen-induced chromosomal aberrations occurring in vivo in epithelial tissues of both the oral cavity and the urinary bladder. The procedure involved taking airdried smears of three sites in the oral cavity of each examined individual. Desquamated urinary bladder cells were collected by centrifugation of freshly voided urine samples. Frequencies of exfoliated cells in these preparations were determined and compared with control values (individuals with no genetic chromosomal instability and no known carcinogene exposure) for these sites. Exforliated cell micronucleus (MEC) frequencies were elevated 5- to 14-fold in samples from the A-T homozygotes. This elevation in MEC frequency occurred for both the oral cavity and urinary bladder. Five out of the seven obligate A-T heterozygotes had an elevated MEC frequency in samples from the oral cavity. In addition, all examined urine samples from A-T heterozygotes contained an elevated percentage of micronucleated cells. These data suggest that this assay is suitable for in vivo monitoring of groups of individuals in which genetically produced chromosomal damage occurs. The possibility of A-T heterozygote detection with this simple procedure is of particular significance, since such individuals are believed to comprise up to 1% of the general population, and have been identified as being at elevated risk for cancer. 相似文献
10.
Regeneration of ventral root axons of the lumbar seventh (L7) segment into the dorsal L7 roots on the opposite side of cat spinal cord was shown by changes in the levels of acetylcholinesterase (AChE) and pseudocholinesterase (PsChE). Low levels of AChE and PsChE were found in control dorsal roots, but when regenerating ventral root fibers entered the dorsal roots, there was a doubling of AChE activity within 2 weeks. Growth appears to start some time after the first week; this is in accord with earlier evidence based on axoplasmic flow of isotope labeled protein in this experimental preparation. The level of AChE activity in the reinnervated dorsal roots increased continually for about 100 days before reaching a plateau at approximately 20 × control levels. The gradual increase and the plateau of AChE activity is in accord with a maturation of the ventral root fibers which had regenerated into the dorsal roots. PsChE in the dorsal roots changes in parallel with AChE in a ratio of 1:10, suggesting that PsChE may in part be localized in the regenerating axons. 相似文献