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I R Johnston

Publications and source records attributed to I R Johnston.

10 recordsLinked to original sources

A DNA-dependent ATPase of calf-thymus.

A DNA-dependent ATPase has been purified from calf thymus. The enzyme hydrolyses ATP and dATP in the presence of heat-denatured DNA. It does not hydrolyse the corresponding nucleoside triphosphates of guanine, uridine and cytosine. The Km values for ATP and dATP are both 0.62 mM. The enzyme requires magnesium or manganese ions. Its sedimentation coefficient is about 4.4 S. The catalytic activity is inhibited by N-ethylmaleimide but is not sensitive to novobiocin and nalidixic acid which are potent inhibitors of bacterial DNA gyrase. In some cases, during purification, chromatographically distinct additional DNA-dependent ATPase activities were detected. Limited proteolysis or covalent modification of the enzyme in the tissues, or during the first steps of its extraction, are probably responsible for the appearance of these chromatographically distinct forms.

Adenosine Triphosphatases

Studies on the purification and properties of a 6.8-S DNA polymerase activity found in calf-thymus DNA polymerase-alpha fraction.

The heterogeneity of calf thymus DNA polymerase-alpha has been further investigated. In particular, an enzyme (enzyme D) which exhibits higher activity on poly(dA) . (dT)10 (A:T = 20:1) compared with that on activated DNA, has been further purified and its properties compared with two other activities of the DNA polymerase-alpha fraction (enzymes A1 and C) which do not show a preference for poly(dA) . (dT)10 over activated DNA. As with A1 and C, enzyme D was shown to have many of the characteristic properties of DNA polymerase-alpha in that it is an acidic protein as judged by its binding to DEAE-cellulose, has a molecular weight of about 140000, does not use a poly (A) . (dT)10 template-initiator complex and is inhibited by N-ethylmaleimide. It exhibits anomalous gel filtration behaviour on Sepharose 6B and it binds relatively weakly to DNA-cellulose compared with DNA polymerase-beta. The extreme sensitivity of enzyme D to inhibtion by N-ethylmaleimide distinguishes it from A1 and C, as does its elution position from a DEAE-cellulose column. On the other hand enzymes C and D are readily inactivated by heating at 45 degrees C unlike enzyme A1. The possible interrelationships of the multiple activities of calf thymus DNA polymerase-alpha are discussed.

Animals

Evidence that DNA polymerase-alpha of calf thymus contains a subunit of molecular weight 155000.

Small samples of the 8-S species of enzymes (A1 and A2) occurring in the DNA polymerase-alpha fraction of calf thymus, have been extensively purified using non-denaturing (normal) polyacrylamide gel electrophoresis. When peak fractions of activity on normal gels were subjected to dodecylsulphate-polyacrylamide gel electrophoresis, a polypeptide at 155000 correlated with polymerase activity. Samples of the 7.3-S (C) enzyme prepared from A2 by treatment with 2.4 M urea or isolated directly without exposure to urea, also showed the presence of a 155000-Mr polypeptide. It is concluded that the 7.3-S (C) enzyme, of previously estimated molecular weight 155000-170000, is a single polypeptide and that the 8-S enzymes A1 and A2 contain an additional subunit of 50000-70000 molecular weight.

Animals

Traffic safety education: panacea, prophylactic or placebo?

Education is one of the strategies available to reduce traffic crashes and the resultant personal injury. It is seen by many as the major strategy for achieving lasting change. However, there is considerable debate as to the effectiveness of traffic safety education programs to date and, in an era when public expenditure is strongly influenced by the results of cost-benefit analyses, the more fundamental traffic safety education programs are under increasing challenge. The literature on effectiveness is indeed confusing. All too often, programs have commenced from a position of blind faith and have been implemented unsystematically, without specific objectives, targets, or evaluative milestones. The problem is compounded by the considerable methodological difficulties which confront the evaluators of long-term programs of behavior change. If traffic safety education is to survive as a viable countermeasure, program planning and execution must become far more scientific and evaluation must become an integral component. There is also an arguable case for the funds for the longer-term behavior change programs to come from the more general public health arena than from the narrower traffic safety field, while the short-term, specific educational programs should compete directly for funds with alternative traffic safety measures available for the given problem.

Accidents, Traffic