Molecular changes in UV-induced and γ-ray-induced mutations in human lymphoblastoid cells

We have characterized the structural changes in the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene of 14 UV-induced, 15 γ-ray-induced and 17 spontaneous mutants of human lymphoblastoid cells selected for 6-thioguanine (6TG) resistance. Southern blot analysis using the full-length HPRT cDNA as a probe revealed that 29% (5/17) of the spontaneous mutants contained detectable alterations in their restriction fragment patterns. Among the 15 mutants induced by γ rays, 7 (47%) had such alterations indicative of large deletions in the HPRT gene. In contrast, all 14 UV-induced mutants exhibited hybridization patterns indistinguishable from those of the wild-type cells. These results suggest that UV is likely to induce point mutations at the HPRT locus on the human chromosome and that the molecular mechanism of UV-induced mutation is quite different from that of ionizing radiation-induced mutation or spontaneous mutation in human cells.     

Salt Adaptation of K+ Channels in the Plasma Membrane of Tobacco Cells in Suspension Culture

The patch-clamp technique was used to study and compare thecharacteristics of cation channels in the plasma membrane ofcultured lines of tobacco (Nicotiana tabacum L. cv. Bright Yellow-2)cells that were unadapted (NaCl-unadapted cells) and adaptedto 50 and 100 mM NaCl (Na50-adapted and Na100-adapted cells).In these three types of tobacco cell, the outward whole-cellcurrent activated by depolarization was dominated mainly bythe activity of the outward rectifying K⁺ channels with a single-channelconductance of 20 pS. The steady-state amplitude of the outwardwhole-cell currents at all the positive potentials examineddecreased in the following order: NaCl-unadapted cells>Na50-adaptedcells>Na100-adapted cells. There were no significant differencesbetween the NaCl-unadapted and the Na50-adapted cells in termsof the ratio of permeabilities of these channels to K⁺ and Na⁺ions. Furthermore, no significant differences in terms of thesingle-channel conductance of these channels were observed amongthe NaCl-unadapted, the Na50-adapted and the Na100-adapted cells.These observations suggest that adaptation to salinity of tobaccocells in suspension results in reduced permeability of the K⁺channels to both K⁺ and Na⁺ ions, without any change in theK⁺/Na⁺ selectivity and single-channel conductance of these channels. ¹Present address: Research Laboratory of Applied Biochemistry,Tanabe Seiyaku Co., Ltd.16-89 Kashima 3-chome, Yodogawaku, Osaka,532 Japan