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Biomedical subjects

Y Sokawa

Publications and source records attributed to Y Sokawa.

At least 73 records · Page 4Linked to original sources
PubMed1983

(2'-5')Oligoadenylate synthetase in pig spleen: isolation and characterization.

A high level of activity of (2'-5')oligoadenylate synthetase (2-5A synthetase) was detected in pig spleens not treated with exogenous interferon. The enzyme recovered from homogenates of pig spleens was partially purified by the use of a combination of ion exchange gels (DEAE-Sephadex and CM-Sepharose) and affinity gels (2',5'-ADP-Sepharose and poly(I):poly(C)-agarose). The specific activity of the final sample was 32,800 nmol AMP polymerized/h/mg protein at 33 degrees C, and the enzyme was able to convert over 80% of ATP into 2-5A after a 24-h incubation; penta-adenylate was the major product (41% of total product). The 2-5A synthetase obtained was eluted from Sephacryl S-200 at around the position of a protein with Mr = 100,000. By using the purified 2-5A synthetase from pig spleens as an antigen, rabbit antiserum to this enzyme was prepared. The antibody bound to protein A-Sepharose absorbed the activity of 2-5A synthetase induced by interferon in pig cells. This result shows that the IFN-induced 2-5A synthetase shares antigenic determinants with the synthetase in spleens. Neither human nor mouse 2-5A synthetase combined with the antibody to porcine 2-5A synthetase. Thus, the antigenic structure of this enzyme is different from species to species.

2',5'-Oligoadenylate Synthetase↗
PubMed1982

Microinjection of interferon and 2',5'-oligoadenylate into mouse L cells and their effects on virus growth.

By means of a new type of microinjection apparatus, which has a micropipette located in a hole through the optical axis of the condenser lens, we injected interferon (IFN) or 2',5'-oligoadenylate (2-5A) into mouse L cells, and observed their antiviral effects on the multiplication of vesicular stomatitis virus (VSV). After injection, cells were infected with VSV, and labeled with [3H]uridine in the presence of actinomycin D. The proportion of cells infected with VSV which carried radioactive virus-RNA was determined by autoradiography. IFN introduced directly into L cells had no effect on the virus growth. This result supports the idea that IFN molecules exert their effect from outside the cell membrane without penetrating into the cytoplasm. 2-5A, on the other hand, was able to inhibit the growth of VSV effectively when injected into L cells. The antiviral effect was dependent on the dose of 2-5A injected, and moreover the effect was transient, since it disappeared completely after 24-h incubation.

Adenine Nucleotides↗
PubMed1980

Induction of 2',5'-oligoadenylate synthetase and interferon in mouse trigeminal ganglia infected with herpes simplex virus.

The activity of 2',5'-oligoadenylate synthetase, which is induced by interferon action, was detected in mouse trigeminal ganglia infected with herpes simplex virus (HSV) type 1. When HSV was inoculated on the left cheek of mice, the virus began to appear in the ipsilateral trigeminal ganglia on day 2, reached its maximum accumulation on day 4, declined thereafter, and was no longer detected in the tissue homogenate after 11 days. After this short acute productive phase, the virus entered into the latent phase of infection. 2',5'-Oligoadenylate synthetase appeard in the trigeminal ganglia shortly after the beginning of virus multiplication; the synthetase activity began to rise on day 3, reached a maxium level on day 4, and then declined. Interferon activity (type I) also appeared in the infected ganglia, but diminished more rapidly than the synthetase. The antibody against HSV in the sera began to appear at the time when the virus and synthetase were declining. From these results it may be hypothesized, although not concluded, that in the acute productive phase of HSV infection in mouse trigeminal ganglia, virus multiplication is suppressed by interferon action, including 2',5'-oligoadenylate synthetase induction.

2',5'-Oligoadenylate Synthetase↗
PubMed1980

Induction of poly(I):poly(C)-binding 50 K protein and 2',5'-oligoadenylate synthetase in interferon-treated mouse L929 cells.

A new protein retained by poly(I):poly(C)-Sepharose was induced together with dsRNA-dependent enzymatic activities, a protein kinase and 2',5'-oligoadenylate synthetase (2,5A synthetase), in interferon-treated mouse L929 cells; it had an apparent molecular weight of 50,000 (50 K) and was not phosphorylated by the protein kinase. The kinetics of the induction of the poly(I):poly(C)-binding 50 K protein were similar to those of dsRNA-dependent protein kinase and 2',5'-oligoadenylate synthetase, and their inductions were all dependent on the interferon dose added, though a relatively higher dose was required for the 50 K protein. When the interferon preparation was heated to 100 degrees C in the presence of sodium dodecyl sulfate, its effect on cells of inducing the activity of 2',5'-oligoadenylate synthetase was preserved completely, indicating that the interferon molecule itself is responsible for the induction of the synthetase. Since the induction of the enzymatic activity was inhibited by addition of either actinomycin D or cycloheximide, it may not be an activation of a latent enzyme but a de novo synthesis of the enzyme.

2',5'-Oligoadenylate Synthetase↗
PubMed1979

2',5'-Oligoadenylate synthetase activity in lymphocytes from normal mouse.

The activity of 2',5'-oligoadenylate synthetase, an enzyme recently discovered in interferon-treated cells, was found in lymphocytes from normal mouse spleen that had received neither exogenous interferon nor its inducers. The oligoadenylate synthesized by lymphocyte cell extracts inhibited protein synthesis in rabbit reticulocyte lysates. The oligomers were composed mainly of trimer and were resistant to digestion by T2 ribonuclease. The level of the enzyme in lymphocytes was about 20 to 30% of that in L929 cells treated with interferon. The activity of the enzyme was further enhanced in lymphocytes in vitro by addition of interferon. The 2',5'-oligoadenylate synthetase was distributed among several lymphoid tissues, but was not detected in cell extracts from brain or liver. The enzyme may play an important role in the regulation of the immune system.

Adenosine Monophosphate↗
PubMed1978

Relaxation effect of chloramphenicol on the stringent control in Escherichia coli.

In 10B601 (rel+) strain possessing a temperature-sensitive valyl-tRNA synthetase, chloramphenicol prevented the formation of guanosine-3'-diphosphate-5'-diphosphate (ppGpp) as well as the stringent control of stable RNA synthesis, under the conditions where the incorporation of valine into protein was still detectable i.e. at the lower restrictive temperatures. On the other hand, the effect of chloramphenicol was not observed at higher restrictive temperatures above 42 degrees C where the incorporation of valine was completely absent. Pretreatment of 10B601 cells with chloramphenicol before transfer to a high restrictive temperature (43.5 degrees C) did retard the onset of accumulation of ppGpp after the shift-up. Duration of the lag period was dependent on the concentration of chloramphenicol added. In parallel with the inability of the cells to accumulate ppGpp, stable RNA synthesis was permitted to continue at that high temperature. These results suggest that chloramphenicol traps aminoacyl-tRNA at the A-sites of ribosomes by damming-up the small flow of aminoacyl-tRNA under the restrictive supply of amino acids. Unchanged tRNA which has been located at the A-site is replaced by the charged one, thus resulting in the suppression of ppGpp formation and in the restoration of stable RNA synthesis.

Biological Transport↗
PubMed1978

Effect of interferon on the cell cycle of BALB/c 3T3 cells.

The effect of interferon on the exponential growth phase of BALB/c 3T3 cells was studied. Although interferon reduced the growth rate and the proportion of cells in both the M and S phases (mitotic index and labelling index), there were no appreciable differences in the duration of these phases between control and interferon-treated cells. Moreover, the shape of the first peak of the fraction of labelled mitoses (FLM) curve was not altered by treatment with interferon, which indicates that the duration of the S and G2 phases was not affected. However, the height of the second peak of the curve in interferon-treated cells was extremely reduced as compared to control culture. These results are compatible with the idea that the suppressive effect of interferon is exerted mainly in the G1 phase (A-state) of the cell cycle of BALB/c 3T3 cells.

Animals↗
PubMed1975

Regulation of stable RNA synthesis and ppGpp levels in growing cells of Escherichia coli.

Under the balanced condition of growth of E. coli cells, no distinct difference is observed in stable RNA and protein synthesis between CP78 (rel+) and CP79 (rel minus), whereas a considerable difference is present in RNA accumulation between NF161 (rel+) and NF162 (rel minus), where NF161 smaller than NF162. The RNA content of NF161 is lower than that of NF162 in four different cultures with different growth rates. These two sets of isogenic pairs of rel+ and rel minus strains are commonly used in the study of rel gene function; however, NF161 is a mutant in the spoT gene whose product may be responsible for the degradation of ppGpp. The basal levels of ppGpp in these four strains growing with three different growth rates were examined: NF161 (rel+ spoT minus) has a much higher content of ppGpp than do other strains. Furthermore, the contents of ppGpp tend to be lower when the above four strains are growing at a faster rate. Thus a close correlation seems to exist between the content of RNA and the basal level of ppGpp under the condition of balanced growth.

Adenosine Triphosphate↗