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Biomedical subjects

A J Clark

Publications and source records attributed to A J Clark.

At least 307 records · Page 17Linked to original sources
PubMed1980

Genetic and physical mapping of recF in Escherichia coli K-12.

Two factor transductional crosses place recF at approximately 82 min on the E. coli chromosome; recF is highly cotransducible with dnaA and gyrB (cou). Transductional analysis with a series of lambda tna specialized transducing phages carrying chromosomal DNA from the tnaA region place recF between dnaA and gyrB. This analysis also indicates that a gene lying in the same region and producing an easily detectable protein (estimated MW of 45 kD) is dnaN and not recF.

Bacteriophage lambda↗
PubMed1980

Analysis of ultraviolet light-induced suppressor mutations in the strain of Escherichia coli K-12 AB1157: an implication for molecular mechanisms of UV mutagenesis.

Genetic analysis of histidine independent (His+) revertants induced by ultraviolet light in the his-4 E. coli strain AB1157 was carried out: 93% carried ochre (UAA) suppressor mutations and 17% carried back mutations to his+ or (intragenic?) suppressors not detectably separable from his-4. Using the specialized transducing lambda psu 2int- phage, which carries supE-supB, it was determined that 87% of the ochre suppressors mapped in the supE-supB region. We were able to deduce that 56% of these affected tRNA1Gln by a CAA leads to TAA change in the tRNA gene while 31% affected tRNA2Gln by TAG- leads to TAA change. Although we were unable to deduce the base substitution of the remaining 13%, the results indicate that most of the suppressor mutations are caused by G:C to A:T transition. These results suggest that the high incidence of supE-supB region suppressor mutation in E. coli by UV would be a reflection of the general feature of UV mutagenesis; i.e. preferential induction of G:C to A:T transition in repairing nonpairing DNA lesions. AI 05371

Bacteriophage lambda↗
PubMed1980

Sequence-specific recombination of plasmid ColE1.

The conjugal transmission of plasmid ColE1 and its derivatives can lead to recA-independent recombination. The recombination is not observed during vegetative growth and takes place specifically between the sequences that are thought to represent the transfer origin of ColE1. This provides genetic evidence for breakage and reunion events during the transmission of ColE1.

Bacteriocin Plasmids↗
PubMed1980

Transposon-mediated conjugational transmission of nonconjugative plasmids.

When coresident with conjugative plasmid pNC21, the nonconjugative deletion F-prime pJC59, which retains the F transfer origin oriT, was transmitted to transconjugants at a frequency comparable to that of pNC21. In addition, pJC59 was transmitted as an independent plasmid, physically separate from pNC21, an example of plasmid donation. In contrast, two plasmids that are derived from F and deleted for the oriT site, pJC61 and pML31, were transmitted at frequencies 10(4) lower than that of pNC21. This low-frequency transmission was associated with the appearance of a new plasmid in the transconjugants. In the case of pML31, we determined that this new plasmid was a recombinant composed of pNC21 and pML31, the latter flanked by two copies of transposable element Tn3. We believe that this recombinant plasmid was formed as an intermediate in the transposition of Tn3 from pNC21 to pML31 and was the vehicle for conjugational transmission of pML31 genes by a process known as plasmid conduction.

Conjugation, Genetic↗
PubMed1979

Diabetic impotence: studies of nocturnal erection during REM sleep.

Nocturnal erections were studied in 30 diabetic patients who complained of impotence and in 11 healthy volunteers. The maximum increase in penile circumference was measured by a penile strain-gauge and recorded on a portable tape-recorder; an external oculogram was recorded simultaneously to identify periods of rapid-eye-movement sleep. The technique gave reproducible results, was acceptable to patients, and was suitable for use in an oridnary hospital ward. Only six diabetics showed a maximum increase in penile circumference of under 15 mm, whereas all but one of the healthy subjects showed maximum increases above this value. Of the six diabetics, five complained of total impotence and had other features of autonomic neuropathy that suggested an organic basis for their impotence. The other patient complained of partial importence, which was probably caused by psychological factors. These findings suggest that the prevalence of organic impotence among diabetics has been overestimated.

Diabetes Complications↗
PubMed1979

W-reactivation of phage lambda in recF, recL, uvrA, and uvrB mutants of E. coli K-12.

W-reactivation is reduced by recF143 and recF144 mutations and is undetectable if a second mutation at either the uvrA or uvrB locus is combined with recF143. The uvrA and uvrB mutations alone block W-reactivation partially. A recL152 mutation also partially blocks W-reactivation by itself. In combination with a uvrB5 mutation, recL125 blocks W-reactivation completely but in combination with recF143, significant residual W-reactivation ability remains. We suggest that the phenomenon of W-reactivation is the result of at least two modes or pathways. The observation that recF143 uvrB5 and recF143 uvrA6 strains permit normal levels of mutagenesis (Kato et al., 1977) but completely block all W-reactivation leads us to suggest further that the mechanism(s) of W-reactivation is at least partly different from that of UV mutagenesis.

Coliphages↗
PubMed1979

Indirect and intragenic suppression of the lexA102 mutation in E. coli B/r.

In Escherichia coli B/r the expression of UV inducible (SOS) functions is under the control of the recA and lexA genes. In this study we have characterized mutants which are altered in their ability to express SOS functions. These mutants were isolated as UV resistant UV nonmutable (Rnm) derivatives of the lexA102 uvrA155 mutant strain WP51. The UV resistance of these Rnm strains is a result of the suppression of lexA102 mediated UV sensitivity. Genetic mapping of rnm mutations shows that the two predominant classes, rnmA and rnmB, map in or very near the lexA and recA genes respectively. rnmA mutations differ from rnmB with regard to recA protein synthesis, rnmA mutations do not restore the ability to express high levels of recA protein after UV treatment whereas rnmB mutations result in constitutive expression of high levels of recA protein. However, both rnmA and rnmB mutant strains inhibit postirradiation DNA degradation. This shows that in rnmA strains, high levels of recA protein are not needed to inhibit postirradiation DNA degradation. The genetic map location and constitutive expression of recA protein synthesis resulting from rnmB mutations suggests that they are operator constitutive mutations of the recA gene. The result that the lexA+ gene is required for the expression of UV mutagenesis in rnmB mutants shows that high levels of recA protein do not circumvent the need for the lexA+ gene product in this process. Thus, while the lexA gene product is required for the induction of recA protein synthesis, lexA must have an additional role in UV induced mutagenesis.

Bacterial Proteins↗
PubMed1979

Deletions generated by the transposon Tn10 in the srl recA region of the Escherichia coli K-12 chromosome.

A negative regulatory gene for the srl operon (srlR) was recognized by the characteristics of an insertion mutation generated by the transposon Tn10 determining tetracycline resistance. This finding is discussed in light of previous hypotheses on the regulation of the srl genes, which mediate metabolism of glucitol (i.e., sorbitol). Mapping showed that the order of genes in this region is: srlR srlD srlC recA alaS. Using two different methods, five mutations of both srl and recA were detected. The phenotype conferred by these mutations, UV sensitivity and extreme recombination deficiency, is characteristic of standard recA point mutants. Three of the mutations were deletions that also removed the genes for tetracycline resistance of the nearby transposon. A fourth mutation ended at a distance from Tn10 sufficient to allow separation of the two by recombination following P1 transduction; our tests did not allow us to conclude whether this mutation was an inversion or a deletion. The fifth mutation was a deletion that seemed to end immediately adjacent to the boundary of Tn10, proximal to recA. Mechanisms for the generation of these srl recA mutations are discussed.

Chromosome Deletion↗
PubMed1978

Assignment of tra cistrons to EcoRI fragments of F sex factor DNA.

We describe here the cloning of single EcoRI fragments from the tra region of F DNA using ColE1::Tn3 as vector. These plasmids, as well as the series of Skurray et al. (Proc. Natl. Acad. Sci. U.S.A. 73:64-68, 1976), have been used to refine the map positions of tra cistrons on the F factor as well as to define a new DNA transfer cistron, traM. The current map of the tra cistrons is presented. None of the known tra cistrons, with the exception of traG, straddles an EcoRI site. The EcoRI site at 82 kilobases splits the traG cistron into two portions, an operator-proximal portion necessary for F pilus synthesis and an operator distal portion involved in conjugation itself. The operon structure of the tra cistrons was reevaluated, and we found that traI is at least partially independent of transcription of the traA to traD operon.

Conjugation, Genetic↗
PubMed1978

Suppressibility of recA, recB, and recC mutations by nonsense suppressors.

Mutations in the recA, recB, and recC genes of Escherichia coli K-12 were surveyed to ascertain whether or not they are suppressed by nonsense suppressors. Several mutations which map in or near the recA gene, but have not been called recA mutations, were also surveyed. An amber recB mutation, recB156, and an amber recC mutation, recC155, were isolated. One recB mutation, recB95, and four recC mutations, recC22, recC38, recC82, and recC83, were found to be suppressed by a UGA suppressor. In addition to the previously isolated amber recA mutation recA99, two other recA mutations, recA52 and recA123, were found to be suppressed by amber suppressor supD32 but not by supE44.

Escherichia coli↗
PubMed1977

Defective excision and postreplication repair of UV-damaged DNA in a recL mutant strain of E. coli K-12.

The mutation recL152 leads to a reduction of excision repair as measured by an increase in the time required to close uvrA uvrB dependent incision breaks, and by a reduction of host cell reactivation ability. Postreplication repair is also delayed when measured in a uvrB5 recL152 double mutant. Such a determination could not be made using the recL152 single mutant because the excision defect led to an accumulation of breaks in the unlabeled high molecular weight DNA to which the labeled DNA synthesized after irradiation must attach in order to achieve normal high molecular weight. Further, the recL gene product seems to be required to rejoin breaks in parental strand DNA which are generated during postreplication repair, since such gaps accumulate in a recL152 uvrB5 double mutant but not in a recL+ uvrB5 single mutant. We have noticed a striking phenotypic similarity between recL152 and polA1 and suggest that recL152 is required for full in vivo activity of DNA polymerase I.

DNA Repair↗