The Hansenula polymorpha PER8 Gene Encodes a Novel Peroxisomal Integral Membrane Protein Involved in Proliferation

We previously described the isolation of mutants of the methylotrophic yeast Hansenula polymorpha that are defective in peroxisome biogenesis. Here, we describe the characterization of one of these mutants, per8, and the cloning of the PER8 gene. In either methanol or methylamine medium, conditions that normally induce the organdies, per8 cells contain no peroxisome-like structures and peroxisomal enzymes are located in the cytosol. The sequence of PER8 predicts that its product (Per8p) is a novel polypeptide of 34 kD, and antibodies against Per8p recognize a protein of 31 kD. Analysis of the primary sequence of Per8p revealed a 39-amino-acid cysteine-rich segment with similarity to the C3HC4 family of zinc-finger motifs. Overexpression of PER8 results in a markedly enhanced increase in peroxisome numbers. We show that Per8p is an integral membrane protein of the peroxisome and that it is concentrated in the membranes of newly formed organdies. We propose that Per8p is a component of the molecular machinery that controls the proliferation of this organelle.

Hansenula polymorpha: An attractive model organism for molecular studies of peroxisome biogenesis and function

In wild-type Hansenula polymorpha the proliferation of peroxisomes is induced by various unconventional carbon- and nitrogen sources. Highest induction levels, up to 80% of the cytoplasmic volume, are observed in cells grown in methanol-limited chemostat cultures. Based on our accumulated experience, we are now able to precisely adjust both the level of peroxisome induction as well as their protein composition by specific adaptations in growth conditions. During the last few years a series of peroxisome-deficient (per) mutants of H. polymorpha have been isolated and characterized. Phenotypically these mutants are characterized by the fact that they are not able to grow on methanol. Three mutant phenotypes were defined on the basis of morphological criteria, namely: (a) mutants completely lacking peroxisomes (Per-; 13 complementation groups); (b) mutants containing few small peroxisomes which are partly impaired in the peroxisomal import of matrix proteins (Pim-; five complementation groups); and (c) mutants with aberrations in the peroxisomal substructure (Pss-; two complementation groups). In addition, several conditional Per-, Pim- and Pss- mutants have been obtained. In all cases the mutant phenotype was shown to be caused by a recessive mutation in one gene. However, we observed that different mutations in one gene may cause different morphological mutant phenotypes. A detailed genetic analysis revealed that several PER genes, essential for peroxisome biogenesis, are tightly linked and organized in a hierarchical fashion. The use of both constitual and conditional per mutants in current and future studies of the molecular mechanisms controlling peroxisome biogenesis and function is discussed.

Hydrologic data for urban studies in the Austin, Texas, metropolitan area, 1981

This technical report includes maps of ground-water data-collection sites, rainfall for specific storms, storm rainfall-runoff for the 1981 water year, monthly water-level measurements of observation wells, water quality data, peak discharge, and daily rainfall summaries for specific gages.Waller Creek Working Grou

The Effect of Prolactin on the Number of Membrane-Associated Particles in Kidney Cells of the Euryhaline Teleost Gasterosteus aculeatus during Transfer from Seawater to Freshwater:A Freeze-Etch Study

The membranes of kidney cells of 3-spined sticklebacks were examined in freeze-etch replicas. The numbers of particles adhering to surfaces and fracture faces of the outer cell membranes and the membranes of the basal labyrinth were determined. The latter membranes probably are the main location of ion-transporting enzyme complexes. The total number of particles per cell in freshwater fish exceeds that of seawater fish by about 50 % for the outer cell membrane, and by almost 200 % for the membranes of the basal labyrinth. After transfer of seawater fish to freshwater, particle numbers increase and their densities approximate freshwater values after 20 h. This rise in particle numbers coincides with the increase of ion-transporting activity of the cells known to take place after transfer to freshwater. The rate of increase of particle densities is enhanced after injection of ovine prolactin. This hormone is known to stimulate Na+/K+-ATPase activity of the basal labyrinth of teleost kidney cells. The results indicate that the particles represent enzyme complexes. The number of particles is probably under hormonal control. The increase in particle densities after transfer to freshwater is accompanied by a rise in the number of nuclear pores, which is noticeable by 10 h. No changes were observed in the density of the particles adhering to the fracture faces of gap junctions

The peroxisomal membrane protein Pex14p of Hansenula polymorpha is phosphorylated in vivo

Hansenula polymorpha Pex14p (HpPex14p) is a component of the peroxisomal membrane essential for peroxisome biogenesis, Here, we show that HpPex14p is phosphorylated in vivo. In wild-type H, polymorpha cells, grown in the presence of [P-32]orthophosphate, the 32P label was incorporated into HpPex14p. Labelled HpPex14p was induced after a shift of cells to methanol-containing media and rapidly disappeared after a shift to glucose medium, which induces specific peroxisome degradation. Alkaline phosphatase treatment of labelled HpPex14p resulted in the release of 32P and a minor shift of the HpPex14p band on Western blots. Phosphoamino acid analysis by two dimensional silica gel thin layer chromatography suggested that the major phosphoamino acid in phosphorylated HpPex14p was acid-labile, (C) 1999 Federation of European Biochemical Societies

The Effect of Prolactin on the Number of Membrane-Associated Particles in Kidney Cells of the Euryhaline Teleost Gasterosteus aculeatus during Transfer from Seawater to Freshwater:A Freeze-Etch Study

The membranes of kidney cells of 3-spined sticklebacks were examined in freeze-etch replicas. The numbers of particles adhering to surfaces and fracture faces of the outer cell membranes and the membranes of the basal labyrinth were determined. The latter membranes probably are the main location of ion-transporting enzyme complexes. The total number of particles per cell in freshwater fish exceeds that of seawater fish by about 50 % for the outer cell membrane, and by almost 200 % for the membranes of the basal labyrinth. After transfer of seawater fish to freshwater, particle numbers increase and their densities approximate freshwater values after 20 h. This rise in particle numbers coincides with the increase of ion-transporting activity of the cells known to take place after transfer to freshwater. The rate of increase of particle densities is enhanced after injection of ovine prolactin. This hormone is known to stimulate Na+/K+-ATPase activity of the basal labyrinth of teleost kidney cells. The results indicate that the particles represent enzyme complexes. The number of particles is probably under hormonal control. The increase in particle densities after transfer to freshwater is accompanied by a rise in the number of nuclear pores, which is noticeable by 10 h. No changes were observed in the density of the particles adhering to the fracture faces of gap junctions

Hydrologic Data for Urban Studies in The Austin, Texas, Metropolitan Area, 1982

The technical report mentions Waller Creek as a main stream in the study area, but a preliminary search provides no results to specific data from the site.Hydrologic investigations of urban watersheds in Texas were begun by the U.S. Geological Survey in 1954. Studies are now in progress in Austin, and Houston. Studies have been completed in the Dallas, Fort Worth, and San Antonio areas. The Geological Survey, in cooperation with the Texas Department of Water Resources, began hydrologic studies in the Austin urban area in 1954. In cooperation with the city of Austin, the program was expanded in 1975 to include additional streamflow and rainfall gaging stations, and the collection of surface water-quality data. In 1978, the program was expanded to include a ground-water resources study of the South Austin metropolitan area in the Balcones Fault Zone.Waller Creek Working Grou