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Biomedical subjects

A J Clark

Publications and source records attributed to A J Clark.

At least 361 records · Page 20Linked to original sources

Biochemical and genetic studies of recombination proficiency in Escherichia coli. I. Enzymatic activity associated with recB+ and recC+ genes.

An adenosine triphosphate-dependent deoxyribonuclease activity has been detected in lysates of recB(+)recC(+) strains. Mutations in recB or recC lead to loss of this activity, suggesting that these two genes determine the nuclease activity. The over-all reaction in crude lysates digests native DNA to nucleoside monophosphates. Complementation between recB21 and recC22 in vivo leads to normal levels of ATP-dependent nuclease activity. No complementation in vitro has been detected. Mutations in a third recombination gene (recA) do not alter significantly the wild-type levels of this nuclease activity.

Adenosine Triphosphate↗

Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec+ revertants caused by indirect suppression of rec- mutations.

All recB(-) and recC(-) mutants of E. coli carry out significant residual genetic recombination, whereas all recA(-) mutants form no recombinants. This observation suggests that an alternative minor pathway of recombination, independent of recB(+) and recC(+) products, may be operative in Escherichia coli. Rec(+) revertants of recB(-)recC(+), recB(+)recC(-), and recB(-) strains of E. coli have been isolated and are shown to fall into at least two major genotypic classes. One class carries revertant mutations which map in or very near the recB and recC genes. In this class an ATP-dependent DNase characteristic of wild type E. coli is restored. The reversions in this class are probably back-mutations or intragenic suppressor mutations. A second class carries revertant mutations which are located far from the recB and recC genes. In this class there is a high level of DNase activity which does not require ATP and is inactive on T4 DNA. Indirect and not informational suppression appears to be responsible for the second class of revertants. The suggestion is made that restoration of recombination by indirect suppression involves an activation or derepression of one or a series of enzymes, which participate in a pathway of recombination, alternative to the recB and recC pathway, but normally of minor importance. The ATP-independent DNase may be one of these enzymes.

Adenosine Triphosphatases↗

Characteristics of some multiply recombination-deficient strains of Escherichia coli.

Strains of Escherichia coli have been made carrying lesions in more than one gene determining recombination. The following genotypes were constructed and verified: recC22 recB21 recA(+), recC22 recB21 recA13, recC22 recB(+)recA13, and recC(+)recB21 recA13. All multiple rec(-) strains carrying recA13 were similar to AB2463, which carries recA13 alone, in their UV sensitivities, recombination deficiencies, and inabilities to induce lambda phage in a lysogen. However, whereas AB2463 shows a high rate of ultraviolet (UV)-induced deoxyribonucleic acid (DNA) breakdown, the multiple rec(-) strains showed the low level characteristic of strains carrying recC22 or recB21 alone. The strain carrying both recC22 and recB21 was similar in all properties to the single mutants, suggesting that both gene products act in the same part of the recombination and UV repair pathways. It is concluded that in a Rec(+) strain, the recA(+) product acts to inhibit DNA breakdown determined by the recC(+) and recB(+) products.

Coliphages↗

Genetic location of certain mutations conferring recombination deficiency in Escherichia coli.

Four mutations conferring recombination deficiency (recA1, recA13, rec-12, and rec-65) have been cotransduced with cysC and pheA; consequently they lie between 53 and 50 on the standard genetic map of Escherichia coli. The four mutations show different degrees of apparent dominance in transient rec(+)/rec(-) zygotes, and the degree of apparent dominance of a particular mutation was shown to be a characteristic of that mutation, not a reflection of other genetic differences in the original rec mutant strain. All four mutations map at similar distances from cysC and pheA and, despite the different degrees of apparent dominance, all may lie in the recA gene.

Arginine↗

Actinomycin D effect on amino acid absorption from rat jejunal loops.

The absorption of amino acids from jejunal loops was suppressed in anesthetized rats treated previously with 1.0 or 1.5 micrograms of actinomycin D per gram of body weight. The absorption of the acidic, neutral, and basic amino acids showed selective differences in response that were relative to the degree of inhibition and to the time interval required to demonstrate actinomycin sensitivity.

Amino Acids↗

Behavior of lambda bacteriophage in a recombination deficienct strain of Escherichia coli.

The behavior of lambda phage in the Rec(-) strain JC-1569 is compared with that in the Rec(+) strain JC-1557. No difference deemed significant was noted in the adsorption rate, latent period, burst size, frequency of lysogenization, and frequency of vegetative phage recombination. The location of the prophage and its mode of insertion in the Rec(-) lysogen of wild-type lambda (lambda(+)) were inferred to be normal from the results of conjugational crosses. Spontaneous and ultraviolet (UV) irradiation induction of lambda(+) were markedly reduced in the Rec(-) lysogen. On the other hand, thermal induction of a mutant lambda (lambdacI857) lysogen of the Rec(-) strain was not reduced and was only slightly affected by UV irradiation. Phage subject to inhibition by lambda immunity failed to multiply in UV-irradiated cells of the Rec(-) lambda(+) lysogen, whereas those not inhibited by this immunity did multiply. It was concluded that the failure of UV to induce lambda(+) in the Rec(-) lysogen was not due to damage to the prophage, but rather to the inability of the irradiated cells to respond by lifting immunity. Preliminary evidence indicates that a single mutation confers recombination deficiency and the inability to lift immunity after UV irradiation. Possible relationships between recombination and the lifting of immunity are enumerated.

Coliphages↗