Hypoxanthine: Guanine phosphoribosyltransferase mutants in Saccharomyces cerevisiae

Summary Yeast mutants lacking activity of the enzyme hypoxanthine: guanine phosphoribosyltransferase (H:GPRT) have been isolated by selecting for resistance to 8-azaguanine in a strain carrying the wild type allele, ade4 ⁺ of the gene coding for amidophosphoribosyltransferase (PRPPAT), the first enzyme of de novo purine synthesis. The mutants excrete purines and are cross-resistant to 8-azaadenine. They are recessive and represent a single complementation group, designated hpt1. Ade4-su, a prototrophic allele of ade4 with reduced activity of PRPPAT, is epistatic to hpt1, suppressing purine excretion and resistance to azaadenine but not resistance to azaguanine. The genotype ade2 hpt1 does not respond to hypoxanthine. Hpt1 complements and is not closely linked to the purine excreting mutants pur1 to pur5. Hpt1 and pur6, a regulatory mutant of PRPPAT, are also unlinked but do not complement, suggesting a protein-protein interaction between H:G-PRT and PRPPAT. Mycophenolic acid (MPA), an inhibitor of de novo guanine nucleotide synthesis, inhibits the growth of hpt1 and hpt1 ⁺. Xanthine allows both genotypes to grow in the presence of MPA whereas guanine only allows growth of hpt1 ⁺. Activity of A-PRT, X-PRT and H:G-PRT is present in hpt ⁺. Hpt1 lacks activity of H:G-PRT but has normal A-PRT and X-PRT.     

Growth and lipolysis of rat adipose tissue: effect of age, body weight, and food intake

The purpose of the present work was to study age- and weight-controlled rats to determine which is the primary factor in reducing the lipolytic response of free fat cells and which has the greater effect on the ratio of fat cells to nonfat cells in adipose tissue. The method for estimating fat cell and nonfat cell numbers is based on the analysis of adipose tissue and fat cell DNA and lipid. In adequately fed rats, epididymal adipocyte hyperplasia is complete between 9 and 14 wk of age. Chronic underfeeding delays, but does not eliminate, normal fat cell hyperplasia and is accompanied by a net loss in the nonfat cell population. During 9-14 wk of age, rat epididymal adipose tissue enlarges mainly through adipocyte hypertrophy. Total fat cells from the epididymal adipose tissue of control rats represent only 20-23% of the total cell population. Chronic underfeeding increases the percentage of fat cells in the fat pad from 23 to 28%. Noradrenaline-stimulated lipolysis is proportional to fat cell numbers but is inhibited when fat cell lipid increases to over 80% of fat pad wet weight. Rat age is apparently not primarily responsible for the decreased noradrenaline-stimulated lipolysis in fat cells of 350-g rats in vitro.     

EXPERIMENTAL OBSERVATIONS ON THE INFLUENCE OF TEMPERATURE,LIGHT, AND SALINITY ON CELL DIVISION OF THE MARINE DIATOM,DETONULA CONFERVACEA (CLEVE) GRAN2

The influence of 116 combinations of temperature (2, 7, 12, 16 C), salinity (5–35‰ at 5‰ intervals) and light (5 levels) on the mean daily cell division rate ( K ) of the Narragansett Bay clone of Detonula confervacea was examined following appropriate preconditioning. Growth did not occur at 16 C, but was excellent (K = 1.2–1.5) under certain combinations of light and salinity at 2, 7, and 12 C, being somewhat better at the 2 highest temperature levels. At 32%, and 1100–1200 ft-c, K increased approximately 2.5 fold from 0.6 to 1.5 between 2 and 12 C. A light-temperature relationship was found which had the general trend of an increased optimal light intensity with increasing temperature. Within the optimal salinity range of 15–30‰, the optimal light intensity was 200–600 ft-c at 2 C, 600–1200 ft-c at 7 C, and 1200–1800 ft-c at 12 C. The light-temperature relationship was most pronounced at 2 and 12 C. At 2 C, K decreased with increasing light intensity, but was independent of this factor at higher temperatures. The optimal salinity range of 15–30‰ was independent of temperature negligible growth occurred at 5‰. In situ and in vitro responses of Detonula confervacea to salinity were in general agreement but its pronounced cryophilic preference in nature (usually reaching maximum abundance below 1 C) contrasts with its excellent growth at 12 C in culture. The experiments suggest that termination of the bloom of Detonula confervacea in Narragansett Bay and elsewhere is not solely temperature-dependent. Temperature does not satisfactorily account for its apparent exclusion from waters contiguous to Narragansett Bay and from other more northerly portions of the northeastern coast of the U.S, or, together with light, for its equally surprising apparent unimportance in Norwegian coastal waters.     

Amino acid uptake in the peripheral nerve of the rat

When rat sciatic nerves were incubated with C¹⁴l-lysine, l- or d-glutamate, or d-l γ-aminoisobutyrate, the labeled compounds penetrated the nerve, and the level of lysine and leucine after 1 hr was higher in the nerve than in the medium. The level increased with time, and at 24 hr glutamate levels also were higher in the nerve than in the medium. Lowering the temperature strongly inhibited uptake, while other conditions such as absence of glucose, absence of sodium, or the presence of cyanide inhibited uptake by nerve less than uptake by brain slices. The uptake against a concentration gradient, and inhibitions of this uptake by metabolic inhibitors and by structural analogs, were interpreted as showing the presence of transport processes for amino acids in peripheral nerves with characteristics similar to such transport processes in the central nervous system.