Biomedical subjects

J W Thatcher

Publications and source records attributed to J W Thatcher.

11 recordsLinked to original sources

PubMed1998

The dynamin-related GTPase, Dnm1p, controls mitochondrial morphology in yeast.

The Saccharomyces cerevisiae Dnm1 protein is structurally related to dynamin, a GTPase required for membrane scission during endocytosis. Here we show that Dnm1p is essential for the maintenance of mitochondrial morphology. Disruption of the DNM1 gene causes the wild-type network of tubular mitochondrial membranes to collapse to one side of the cell but does not affect the morphology or distribution of other cytoplasmic organelles. Dnm1 proteins containing point mutations in the predicted GTP-binding domain or completely lacking the GTP-binding domain fail to rescue mitochondrial morphology defects in a dnm1 mutant and induce dominant mitochondrial morphology defects in wild-type cells. Indirect immunofluorescence reveals that Dnm1p is distributed in punctate structures at the cell cortex that colocalize with the mitochondrial compartment. These Dnm1p-containing structures remain associated with the spherical mitochondria found in an mdm10 mutant strain. In addition, a portion of Dnm1p cofractionates with mitochondrial membranes during differential sedimentation and sucrose gradient fractionation of wild-type cells. Our results demonstrate that Dnm1p is required for the cortical distribution of the mitochondrial network in yeast, a novel function for a dynamin-related protein.

Actins↗

PubMed1998

Mitochondrial fusion in yeast requires the transmembrane GTPase Fzo1p.

Membrane fusion is required to establish the morphology and cellular distribution of the mitochondrial compartment. In Drosophila, mutations in the fuzzy onions (fzo) GTPase block a developmentally regulated mitochondrial fusion event during spermatogenesis. Here we report that the yeast orthologue of fuzzy onions, Fzo1p, plays a direct and conserved role in mitochondrial fusion. A conditional fzo1 mutation causes the mitochondrial reticulum to fragment and blocks mitochondrial fusion during yeast mating. Fzo1p is a mitochondrial integral membrane protein with its GTPase domain exposed to the cytoplasm. Point mutations that alter conserved residues in the GTPase domain do not affect Fzo1p localization but disrupt mitochondrial fusion. Suborganellar fractionation suggests that Fzo1p spans the outer and is tightly associated with the inner mitochondrial membrane. This topology may be required to coordinate the behavior of the two mitochondrial membranes during the fusion reaction. We propose that the fuzzy onions family of transmembrane GTPases act as molecular switches to regulate a key step in mitochondrial membrane docking and/or fusion.

Cytoplasm↗

PubMed1998

Marginal fitness contributions of nonessential genes in yeast.

Analysis of the complete genome sequence of Saccharomyces cerevisiae confirms and extends earlier evidence that a majority of yeast genes are not essential, at least under laboratory conditions. Many fail to yield a discernible mutant phenotype even when disrupted. Genes not subject to natural selection would accumulate inactivating mutations, so these "cryptic" genes must have functions that are overlooked by the standard methods of yeast genetics. Two explanations seem possible: (i) They have important functions only in environments not yet duplicated in the laboratory and would have conditional phenotypes if tested appropriately. (ii) They make small, but significant, contributions to fitness even under routine growth conditions, but the effects are not large enough to be detected by conventional methods. We have tested the second "marginal benefit" hypothesis by measuring the fitnesses of a random collection of disruption mutants in direct competition with their wild-type progenitor. A substantial majority of mutant strains that lack obvious defects nevertheless are at a significant selective disadvantage just growing on rich medium under normal conditions. This result has important implications for efforts to understand the functions of novel genes revealed by sequencing projects.

Evolution, Molecular↗

PubMed1997

Morphogenesis of denticles and hairs in Drosophila embryos: involvement of actin-associated proteins that also affect adult structures.

We sought components that function in morphogenetic events downstream from the segmentation pathway in Drosophila embryos, so we examined mutations that affect development of adult hairs and/or bristles to identify a subset that also affect hairs and denticles on the cuticle of first instar larvae. Mutations at 4 of 23 loci surveyed cause distinct abnormalities in these larval structures, and two other loci have more subtle, variable effects. In particular, forked and singed mutants produce complex, allele-specific phenotypes. These loci encode actin-associated proteins and, consistent with that information, mutations cause abnormalities in actin bundles that support nascent hairs and denticles in stage 14-16 embryos. We suggest that interactions between these and other actin-associated proteins are important in generating the diverse shapes of the cuticular specializations seen in both larvae and adults.

Actins↗

PubMed1975

The effect of diazepam and fentanyl on mental, psychomotor and electroencephalographic functions and their rate of recovery.

Ten healthy male subjects received diazepam (10 or 20 mg), fentanyl (0.1 or 0.2 mg) or a placebo intravenously at weekly intervals according to a latin square design. They were tested on a battery of psychological and electroencephalographic tests at 0.5, 2, 6, and 8 hrs following injection. Fentanyl had little effect on memory while diazepam reduced the ability to learn without increasing forgetting of material alread acquired. By the 2nd hour post injection, only the low dose of fentanyl had no residual effect. Recovery was complete by the 6th hour for all treatments according to the psychological tests except for the lagging effect of high dose of diazepam on memory. The electroencephalographic effects of diazepam persisted beyond the end of the testing sessions while those of the high dose of fentanyl recovered by the 8th hour. Thus in the dosages tested, diazepam had more intense and prolonged effects than fentanyl.

Adult↗

PubMed1974