PubMed HealthSearch

Biomedical subjects

C W Magill

Publications and source records attributed to C W Magill.

6 recordsLinked to original sources

Role of purine base excretion in regulation of purine pools.

Wild type and mutant strains of Neurospora crassa excrete hypoxanthine, xanthine, and uric acid, but not adenine or inosine, when exogenous adenine is added to growing cultures. No detectable excretion occurs in the absence of adenine. The de novo pathway of purine biosynthesis was found to influence the excretion, in that a metabolic block immediately prior to IMP significantly decreased the excretion, while a metabolic block immediately after IMP significantly increased the excretion over that of wild type. The purine catabolic pathway, which is sensitive to ammonia regulation, was found to be a key determinant in the amount and type of excretion. Recently, it was suggested that hypoxanthine accumulation is the result of a mechanism to regulate the adenylate pool size (Leung and Schramm, 1978). In this report, the possibility that hypoxanthine excretion controls adenylate and guanylate pool sizes is discussed and the role of the purine nucleotide cycle in hypoxanthine excretion is examined.

Adenine

Depression of uracil uptake by ammonium in Neurospora crassa.

The mechanism of uracil uptake and one aspect of its regulation were studied in germinated conidia of Neurospora crassa. Uracil was found to be taken up by a transport mechanism that did not exhibit Michaelis-Menten kinetics. Rather, the kinetic patterns indicated two separate systems or a single transport mechanism with negative cooperativity. Cytosine and thymine inhibited uracil uptake, but uridine did not. The mutant strain uc-5-pyr-1, which failed to transport uracil, was used in reversion studies and to map the uc-5 locus. Spontaneous reversion rates at the uc-5 locus were found to be approximately 2 x 10(-8), indicating that the uc-5 lesion results from a single mutation. Loss of the uracil transport function through a single mutation favors the model of a single transport mechanism with negative cooperativity. Uracil uptake was significantly decreased in the presence of NH 4+, and evidence is presented for repression by NH4+ of a uracil transport system. Growth rates of pyrimidine-requiring and wild-type strains measured in the presence and absence of NH4+, with uracil as the pyrimidine supplement, showed that NH4+ decreased the growth rates of the pyrimidine-requiring strains significantly, while having no effect on wild-type growth rates.

Ammonia

Regulation of hypoxanthine transport in Neurospora crassa.

Hypoxanthine uptake and hypoxanthine phosphoribosyltransferase activity (EC 2.4.2.8) were determined in germinated conidia from the adenine auxotrophic strains ad-1 and ad-8 and the double mutant strain ad-1 ad-8. The mutant strain ad-1 appears to lack aminoimidazolecarboximide ribonucleotide formyltransferase (EC 2.1.2.3) or inosine 5'monophosphate cyclohydrolase (EC 3.5.1.10) activities, or both, whereas the ad-8 strain lacks adenylosuccinate synthase activity (EC 6.3.4.4). Normal (or wild-type) hypoxanthine transport capacity was found to the ad-1 conidia, whereas the ad-8 strains failed to take up any hypoxanthine. The double mutant strains showed intermediate transport capacities. Similar results were obtained for hypoxanthine phosphoribosyl-transferase activity assayed in germinated conidia. The ad-1 strain showed greatest activity, the ad-8 strain showed the least activity, and the double mutant strain showed intermediate activity levels. Ion-exchange chromatography of the growth media revealed that in the presence of NH+/4, the ad-8 strain excreted hypoxanthine or inosine, the ad-1 strain did not excrete any purines, and the ad-1 ad-8 double mutant strain excreted uric acid. In the absence of NH+/4, none of the strains excreted any detectable purine compounds.

Biological Transport, Active

Heterokaryosis and parasexuality in Pyricularia oryzae Cavara.

Auxotrophs in three different races of Pyricularia oryzae were obtained following ultraviolet mutagenesis, and tested for complementarity on minimal medium. Prototrophic growth resulted between combinations of different auxotrophs of single strains, but not between strains. The growth did not result form crossfeeding, but required at least transitory heterokaryosis. Putative diploids, i.e., large prototrophic conidia, were readily isolated from the heterokaryons. In addition, conidia with nearly every possible combination of parental markers were found, showing that P. oryzae does indeed complete the parasexual cycle. The degree of genetic recombination found here may help to explain the extreme variability of this organism.

Cell Nucleus

Purine base transport in Neurospora crassa.

Observations presented in this paper point to the presence of dual transport mechanisms for the base adenine in Neurospora crassa. Competition for transport, as well as growth inhibition studies using an ad-1 auxotroph, show that the purine bases adenine, guanine, and hypoxanthine share at least one transport mechanism which is insensitive to adenosine, cytosine, and a variety of other purine base analogues. On the other hand, uptake of adenine by an ad-8 mutant strain unable to transport [8-14C]hypoxanthine at any concentration was not inhibited by guanine or hypoxanthine. This observation demonstrates the existence of an adenine-specific transport system which was also found to be insensitive to inhibition by other purine base analogues, adenosine or cytosine. Recombination analysis of ad-8 by wild-type crosses showed that the inability to transport [8-14C]hypoxanthine was a consequence of the ad-8 lesion or a closely linked mutation. Saturation plots of each system gave intermediary plateaus and nonlinear reciprocal plots which, based on comparison with pure enzyme kinetic analysis, suggest that either each system consists of two or more uptake systems, at least one of which exhibits cooperativity, or that each system is a single uptake mechanism which possesses more than two binding sites where the relative affinity for the purine base first decreases and then increases as the sites are filled.

Adenine