We have studied the comparative behavior of human DNA polymerases alpha and beta on a polynucleotide template of dT100 with dA15 covalently attached at the 3' end to serve as primer when defined numbers of pyrimidine dimers are introduced by UV (254 nm) irradiation. We have obtained the surprising result that with both alpha and beta polymerases the incorporation of labelled dATP is enhanced when the template has been irradiated (maximum value at 1000 J/m2 UV incident dose). In the presence of Mn2+, DNA polymerase beta produces a product size corresponding to full copying of the template whether irradiated or not. In marked contrast DNA polymerase alpha produces only short products on unirradiated strands but full copying of irradiated templates. Evidently both polymerases utilize a much larger fraction of the template pool following UV irradiation.

DNA Polymerase I↗

PubMedDate not supplied

Effects of insulin exposure upon the metabolism of rat aortic media: influence of hydrostatic forces.

When added in vitro, insulin exerts little or no effect on the metabolism of arterial media. However, indirect evidence suggests that an in vivo exposure of arterial tissue to insulin stimulates its metabolism. To evaluate the possibility that hemodynamic forces existing only in vivo may account for this difference, we studied the effect of brief (9-minute) exposures to insulin of isolated rat thoracic aorta perfused in situ at low (34 mm Hg = 4.6 kPa) and high (70 mm Hg = 9.4 kPa) static pressures. After perfusion, incorporation of glucose-derived 14C into CO2 and lipids, and production of lactate by incubated aortic tissue, were measured. At low perfusion pressure, insulin did not affect the metabolism of the media but strongly stimulated all aspects of the glucose metabolism of the adventitia. At high pressure, insulin again stimulated the adventitia but now also significantly stimulated (+66%) the synthesis of glucose-derived lipids by the media; production of CO2 and lactate by the media did not increase significantly. These results indicate that insulin directly stimulates at least one pathway of glucose metabolism by the media; and that hemodynamic forces are an important factor in determining the response of aortic media to insulin.

Animals↗

PubMedDate not supplied

[Increased tRNA concentration in yeast containing mutant termination translation factors eRF1 and eRF3].

Earlier we have characterized strains bearing mutations in essential genes SUP45 and SUP35 of yeast S. cerevisiae, encoding translation termination factors eRF1 and eRF3 respectively. In the present work nonsense-mutants on genes SUP45 and SUP35 have been compared by a level of eight tRNA: tRNATyr, tRNAGln, tRNATrp, tRNALeu and tRNAArg (previously described as potentially suppressor tRNA), and also tRNAPro, tRNAHis and tRNAGly. We have not revealed preferable increase in amount of natural suppressor tRNA. The majority of the investigated mutations leads to increase in a level of all investigated tRNA. The mechanisms providing viability of nonsense-mutants on essential genes SUP45 and SUP35 are discussed.

Alleles↗

PubMedDate not supplied