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Biomedical subjects

P Karran

Publications and source records attributed to P Karran.

86 records · Page 5Linked to original sources
PubMed1978

Enzymatic excision of free hypoxanthine from polydeoxynucleotides and DNA containing deoxyinosine monophosphate residues.

A DNA glycosylase that releases free hypoxanthine by selective cleavage of dIMP residues in polydeoxynucleotides and DNA containing this nonconventional nucleotide is present in Escherichia coli cell extracts. The partly purified enzyme, termed hypoxanthine-DNA glycosylase, does not require divalent cations or phosphate for activity, and it acts more efficiently on double-stranded than on single-stranded substrates. The enzyme has properties different from either uracil-DNA glycosylase or 3-methyladenine-DNA glycosylase and is present at normal levels in E. coli mutants deficient in either of the latter two DNA glycosylases.

Bacillus subtilis↗
PubMed1978

Repair of depurinated DNA in vitro by enzymes purified from human lymphoblasts.

Alkali-labile lesions introduced into T7 DNA by treatment with methyl methanesulfonate were removed and the DNA was repaired by incubation with DNA polymerase alpha and nuclease from a human lymphoblastoid line followed by the addition of DNA ligase. The nuclease preparation contains both apurinic endonuclease and 5'-3' exonuclease activities. Dinucleotides appear to be the first product of exonuclease action. Repair of methyl methanesulfonate-induced damage can occur by the insertion of only a few nucleotides per lesion as in vivo.

Burkitt Lymphoma↗
PubMed1977

Intermediates in excision repair by human cells: use of S1 nuclease and benzoylated naphthoylated cellulose to reveal single-strand breaks.

Treatment of human lymphoid cells with methyl methanesulfonate (MMS) results in discontinuities in cellular DNA. Discontinuities can be detected by sedimentation through neutral sucrose gradients after digestion with S1 nuclease on top of the gradient and by benzoylated naphthoylated diethylaminoethylcellulose (BND-cellulose) chromatography. DNA with discontinuities is sensitive to shear which produces single-stranded regions and causes the DNA to adhere to BND-cellulose. The shear-sensitive and S1-sensitive sites produced by MMS treatment are probably identical. Treatment of cells with acetoxyacetylaminofluorene (AAAF) does not result in detectable discontinuities either at doses which produce the same number of adducts as observed after MMS treatment or at doses which inhibit DNA excision repair. Addition of aromatic adducts to DNA can also result in adherence to BND-cellulose but such DNA is not S1 sensitive. We conclude that MMS- and AAAF-induced damage are repaired by different pathways in human cells and that DNA with discontinuities is a readily detectable intermediate only in the repair of MMS damage by the apurinic pathway.

Acetoxyacetylaminofluorene↗
PubMed1977

Developmental decline in DNA repair in neural retina cells of chick embryos. Persistent deficiency of repair competence in a cell line derived from late embryos.

Neural retinas of 6-day-old chick embryos synthesize DNA and are able to carry out DNA excision repair. However, in contrast to the situation in human cells, the maximum rate of repair induced by N-acetoxy acetylaminofluorene (AAAF) is no greater than that induced by methyl methanesulfonate (MMS). With advancing differentiation of the retina in the embryo, cell multiplication and DNA synthesis decline and cease, and concurrently the cells lose the ability to carry out DNA excision repair. Thus, in 15-16-day embryos, in which the level of DNA synthesis is very low, DNA repair is barely detectable. If retinas from 14-day embryos are dissociated with trypsin and the cell suspension is plated in growth- promoting medium, DNA synthesis is reinitiated; however, in these cultures there is no detectable repair of MMS-induced damage, and only low levels of repair are observed after treatment with AAAF. A cell line was produced, by repeated passaging of these cultures, in which the cell population reached a steady state of DNA replication. However, the cell population remained deficient in the ability to repair MMS-induced damage. This cell line most likely predominantly comprises cells of retino-glial origin. Possible correlations between deficiency in DNA repair mechanisms in replicating cells and carcinogenesis in neural tissues are discussed.

Acetoxyacetylaminofluorene↗
PubMed1976

DNA excision-repair deficiency of human peripheral blood lymphocytes treated with chemical carcinogens.

Human peripheral blood lymphocytes stimulated with concanavalin A for 72 hr have a 10-fold greater capacity to repair DNA damage induced by N-acetoxy-2-acetylaminofluorene than do unstimulated cells. The increased capacity of concanavalin A-activated cells to repair DNA is not observed after 24 hr in culture, a time at which stimulated cells have not begun to synthesize DNA. The maximum rate of repair synthesis obtained after treatment of stimulated cells with the "large patch"-inducing agent, N-acetoxy-2-acetylaminofluorene, is twice that obtained with methyl methanesulfonate, an agent inducing "small patch" repair. The difference between the maximum rates obtained with N-acetoxy-2-acetylaminofluorene and methyl methanesulfonate is 6-fold in a human lymphoblastoid line. Unstimulated lymphocytes show almost identical rates of repair after treatment with either N-acetoxy-2-acetylaminofluorene or methyl methanesulfonate. There is close correlation between the rate of N-acetoxy-2-acetylaminofluorene-induced repair synthesis and the loss of acetylaminofluorene adducts from DNA. Treatment of lymphocytes with methyl methanesulfonate leads to degradation of cellular DNA with the production of single-stranded regions. Such degradation is not observed with N-acetoxy-2-acetylaminofluroene. We conclude that the rate of excision repair is a function of the capacity of cells for DNA synthesis and that lymphocytes that do not synthesize DNA have a limited repair capacity and cannot be used to distinguish between large and small patch repair.

Acetoxyacetylaminofluorene↗
PubMedDate not supplied

Regulation of O6-methylguanine-DNA methyltransferase expression in the Burkitt's lymphoma cell line Raji.

We have investigated the expression of the DNA-repair enzyme O6-methylguanine-DNA methyltransferase in the Burkitt's lymphoma cell line Raji. An existing mutant Raji cell line which lacks thymidine kinase activity had previously been shown to be Mex- and to no longer express O6-methylguanine-DNA methyltransferase. We report here that in addition to the methyltransferase and thymidine kinase, a third enzyme with an unrelated function, galactokinase, is also not expressed in Raji cells. The control of thymidine kinase expression is post-transcriptional and it is possible that galactokinase and methyltransferase can share a common post-transcriptional regulation with thymidine kinase.

Blotting, Southern↗
PubMedDate not supplied

Mismatch binding proteins and tolerance to alkylating agents in human cells.

The Mex- (Mer-) phenotype of human cells is characterised by a sensitivity to agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and N-methyl-N-nitrosourea (MNU). The hypersensitivity of Mex- cells is a consequence of their failure to express the DNA-repair enzyme m6-Gua-DNA methyltransferase. Resistance to MNNG and MNU may be acquired by Mex- cells either by reexpression of a methyltransferase function or by an ill-defined process of tolerance in which the cytotoxic potential of m6-Gua is circumvented without the altered base being removed from DNA. It has been suggested that tolerance might involve an altered mismatch correcting function. We have investigated proteins which recognise and bind specifically to DNA fragments containing single-base mismatches. Cell-free extracts of a Burkitt's lymphoma cell line (Raji) contain two such mismatch binding activities. Neither protein appears to have a high affinity for m6-Gua-containing base pairs. The data indicate that m6-Gua-containing base pairs might be poor substrates for mismatch repair processes in human cells.

Alkylating Agents↗