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Biomedical subjects

M Sekiguchi

Publications and source records attributed to M Sekiguchi.

At least 703 records · Page 39Linked to original sources
PubMed1975

Excision of pyrimidine dimers in normal and T4-infected Escherichia coli: effect of polA and other mutations.

Strains carrying both polA1 and recBts1 mutations, which are defective in DNA polymerase I and have thermolabile exonuclease V (the recBC enzyme), are viable at 30 degrees C but not at 42 degrees C. These mutants exhibit almost normal rate of dimer excision in vivo even at the restrictive temperature. Similar results were obtained with other polA minus strains. We have also investigated effect of host and phage mutations on excision of dimers in T4-infected cells. Only a small amount of dimer is excised in T4v-1-infected cells whereas an extensive and selective release of dimers takes place in T4D-infected cells. Other phage mutations, including mutations in gene 43 and gene 30, do not affect excision of dimers in infected cells.

Coliphages↗
PubMed1975

Enzymic mechanism of excision-repair in T4-infected cells.

Excision of pyrimidine dimers from ultraviolet-irradiated DNA in a cell-free system of Escherichia coli infected with bacteriophage T4 consists of two different steps, one to induce a single-strand break at a point close to a pyrimidine dimer and the other to release dimer-containing nucleotide from the DNA. The enzymes responsible for these steps were isolated and the reactions were characterized; T4 endonuclease V introduces a break at the 5' side of a dimer and 5' in equilibrium 3' exonucleases, which are also induced by T4, act at the break to excise dimer-containing nucleotides. We isolated temperature-dependent v mutants, which exhibit increased sensitivity to UV at 42 degrees C but not at 30 degrees C, and found that the mutants induce temperature-sensitive T4 endonuclease V, indicating that the v gene of T4 is indeed the structural gene for T4 endonuclease V and that the enzyme is responsible for the first step of excision-repair. A possible mechanism of excision-repair in T4-infected cells is discussed.

Coliphages↗
PubMed1975

Restoration of ultraviolet-induced unscheduled DNA synthesis of xeroderma pigmentosum cells by the concomitant treatment with bacteriophage T4 endonuclease V and HVJ (Sendai virus).

Ultraviolet (UV)-induced unscheduled DNA synthesis of xeroderma pigmentosum cells, belonging to complementation groups A, B, C, D, and E, was restored to the normal level by concomitant treatment of the cells with T4 endonuclease V and UV-inactivated HVJ (Sendai virus). The present results suggest that (1) T4 endonuclease molecules were inserted effectively into the cells by the interaction of HVJ with the cell membranes, (2) the enzyme was functional on human chromosomal DNA which had been damaged by UV irradiation in the viable cells, (3) all the studied groups of xeroderma pigmentosum ("variant" was not tested) were defective in the first step (incision) of excision repair.

Cells, Cultured↗
PubMed1975

Biochemical studies on radiation-sensitive mutations in bacteriophage T4-1.

Mutants of bacteriophage T4 which exhibit increased sensitivity to ultraviolet radiation were isolated and their abilities to induce an enzyme system to excise pyrimidine dimers were examined. Among 16 mutants isolated, 12 mutants were found to be defective in inducing T4 endonuclease V, which catalyzes the formation of dimer-specific breaks in ultraviolet-irradiated DNA. A leaky v mutant, which exhibits intermediate ultraviolet sensitivity, was also isolated; the mutant induces a low level of endonuclease V and excises only a small amount of kimers in vitro. Three other mutants, as well as x and y mutants, were able to induce both T4 endonuclease V and dimer-excision enzyme (5'yields 3' exonuclease).

Cell Survival↗
PubMed1974

Biochemical studies on the chi mutation of bacteriophage T4: differential inhibition of chi+ and chi DNA synthesis by mitomycin C.

Biochemical studies were carried out to determine the effect of chi mutation on T4 DNA synthesis. The rate and final extent of DNA synthesis are almost the same with T4D- and T4chi-infected cells, although the burst size of T4chi is about one-sixth that of the wild type. The DNA synthesis of T4chi-infected cells is more readily inhibited by mitomycin C than is that of T4 wild type. When mitomycin C was added during active phage growth, DNA synthesis of T4chi halted almost immediately. T4 DNA polymerases isolated from chi(+)- and chi-infected cells, however, exhibit no difference with regard to their sensitivities to mitomycin C, priming activities with alkylated or ultraviolet light-irradiated templates and other enzymatic properties.

Alkanesulfonates↗
PubMed1971

Infection of actinomycin-permeable mutants of Escherichia coli with urea-disrupted bacteriophage.

Intact cells of actinomycin-permeable mutants of Escherichia coli could be infected with urea-disrupted phage T4 (designated as T4pi). The parental strains and the revertants, which are impermeable to actinomycin, were not susceptible to T4pi unless they had been treated with agents which altered their permeability. The permeable mutants developed competence for pi infection during the growth cycle; cells in the early stationary phase produced 10- to 100-fold more plaques on plating with T4pi than did exponentially growing cells. Colistin (polymyxin E) was effective in converting noncompetent cells of either permeable or nonpermeable strains to the competent state. Treatment with lysozyme resulted in a considerable increase in susceptibility to T4pi of permeable mutants but not of nonpermeable cells. It appears that development of competence for pi infection is mainly due to alterations in the permeability barriers of the cell.

Cell Wall↗