The HYP2 gene of Saccharomyces cerevisiae is essential for aerobic growth: characterization of different isoforms of the hypusine-containing protein Hyp2p and analysis of gene disruption mutants

In Saccharomyces cerevisiae, hypusine-containing proteins are encoded by two closely related genes, HYP1 and HYP2, which are regulated reciprocally by oxygen and heme. We have purified the aerobically expressed hypusine-containing proteins from yeast. The three proteins detected (two isoforms, which differ in their pI values, and a degradation product thereof, lacking the N-terminal 10 amino acid residues) are all encoded by HYP2. The N-terminus of both isoforms is formed by acetylation of a serine residue after cleavage of the first methionine. Cells mutant for hyp2 are unable to grow aerobically. However, under anaerobic conditions these mutants display no obvious phenotype, presumably because the strictly anaerobically expressed HYPI gene product (Hyp1p) is present. This implies that Hyp1p and Hyp2p fulfill very similar functions. In fact, Hyp1p can substitute for Hyp2p under aerobic conditions, when expressed under the control of the GAL1 promoter in hyp2 mutant cells.Abbreviations HYP1 and HYP2 S. cerevisiae genes encoding hypusine-containing protein Hyplp and Hyp2p, respectively     

Über die chromotropen Zellen der Nebenniere vom Wasserfrosch

Zusammenfassung Aus dieser neuerlichen Untersuchung der zuerst von Stilling beschriebenen Sommerzellen ergibt sich somit als einzige Übereinstimmung in den sonst widerspruchsvollen Angaben des Schrifttums, daß sie in ihrem Vorkommen nicht auf den Sommer beschränkt sind, weshalb sie von mir nach ihrer färberischen Eigentümlichkeit als chromotrope Zellen bezeichnet werden. Da ihr isoelektrischer Punkt bei etwa p_H 5 liegt, verhalten sie sich gegenüber Farbstoffen nicht ausschließlich (acido-) oxyphil, doch sind sie auch keine Mastzellen oder überhaupt während der Entwicklung veränderte Wanderzellen, sondern eine besondere Art autochthon entstandener Nebennieren-Epithelzellen, deren Körnchen saure Polysaccharide enthalten. Ihre funktionelle Bedeutung muß erst geklärt werden. Dabei ist es besonders bemerkenswert, daß sich das Vorkommen dieser Zellen auf Rana esculenta und eine Abart von ihr sowie exotische Verwandte beschränkt, während sie bei Rana temporaria und deren nächsten Verwandten immer fehlen. Die gegenteiligen Angaben der Literatur beruhen wahrscheinlich teilweise auf unzutreffender Bestimmung der Art und im übrigen wohl auf Verwechslung mit gekörnten Wanderzellen, was besonders bei der Entwicklung zu falschen Vorstellungen führen kann.Herrn Professor Alfred Kohn in dankbarer Erinnerung gewidmet.     

15N nmr spectroscopy. I. Polysarcosine and related sequence polypeptides

The natural abundance ¹⁵N nmr spectra of linear polysarcosine (DP = 35) has been recorded in Me₂SO and H₂O solution. Because of cis/trans isomerization at the peptide bond, a broad signal with several splittings was observed. These splittings appear to reflect the influence of three peptide bonds on a single N atom. The ¹⁵N signals from the sequence polypeptides (β-Ala-Sar-Gly)_n and (β-Ala-Sar-D ,L -Ala)_n also show a cis/trans splitting, as well as chemical shifts which are dependent on the peptide sequence. The tertiary nitrogen of the sarcosyl residue has a T₁ relaxation time which is longer than the T₁ for secondary nitrogens of the other amino acids. The nuclear Overhauser effect is also discussed.     

The cellular mechanism for the formation of lamellar bodies in type-II pneumocytes in rat lungs]

Pneumocyte type II cells from lungs of native rats and of rats that inspired a hypoxic mixture of gases were investigated by transmission electron microscopy. In cells of experimental rats, membrane structures were found that well compare with lamellar bodies. Experimental results and analysis of literature allowed to put forward a hypothesis about the cell mechanism of formation of lamellar bodies from the spiral twisted membranes of the endoplasmic reticulum.     

Improving performance of docking-based virtual screening by structural filtration

In the current study an innovative method of structural filtration of docked ligand poses is introduced and applied to improve the virtual screening results. The structural filter is defined by a protein-specific set of interactions that are a) structurally conserved in available structures of a particular protein with its bound ligands, and b) that can be viewed as playing the crucial role in protein-ligand binding. The concept was evaluated on a set of 10 diverse proteins, for which the corresponding structural filters were developed and applied to the results of virtual screening obtained with the Lead Finder software. The application of structural filtration resulted in a considerable improvement of the enrichment factor ranging from several folds to hundreds folds depending on the protein target. It appeared that the structural filtration had effectively repaired the deficiencies of the scoring functions that used to overestimate decoy binding, resulting into a considerably lower false positive rate. In addition, the structural filters were also effective in dealing with some deficiencies of the protein structure models that would lead to false negative predictions otherwise. The ability of structural filtration to recover relatively small but specifically bound molecules creates promises for the application of this technology in the fragment-based drug discovery.     

Glycosylated equine prolactin and its carbohydrate moiety

Glycosylated equine prolactin (G-ePRL) and nonglycosylated ePRL were purified to homogeneity from side fractions obtained during isolation of LH/FSH from horse pituitaries. Both PRL forms were isolated together in high yield by the isolation procedure used for glycosylated porcine PRL/(G-pPRL) and pPRL, involving acetone extraction/precipitation, NaCl and isoelectric precipitation, and gel filtration. Purification of G-ePRL required additional Con A chromatography. The N-terminal amino acid sequencing for 32 cycles of G-ePRL and ePRL resulted in sequences identical to the known primary structure of ePRL. Based on MALDI mass spectrometry analysis and SDS-PAGE mobilities,G-ePRL and ePRL had estimated molecular weights of 25,000 and 23,000 Da, respectively. G-ePRL displayed only 60% of the immunoreactivity of ePRL in homologous radioimmunoassay. Using the Nb2 lymphoma cell bioassay, ePRL was found to have about l/30th the mitogenic activity of bovine PRL; G-ePRL was approximately l/10th as active as ePRL. Glycosylation of G-ePRL at Asn³¹ was confirmed by isolation and sequence analysis of an enzymatically derived G-ePRL glycopeptide spanning residues 29–37. Monosaccharide compositions of intact G-ePRL and this glycopeptide were very similar (Man₃, GlcNAc₂, GalNAc₁, Fuc_0.6, Gal_0.2, NeuAc_0.15) and resembled that of G-pPRL. The glycopeptide contained one sulfate residue as determined by ion chromatography after acid hydrolysis, indicating the presence of a sulfated monosaccharide. Comparative carbohydrate analysis of G-ePRL and other G-PRL preparations suggests that the functionally significant Asn³¹ carbohydrate unit is a fucosylated complex mono- and/or biantennary oligosaccharide terminating with a sulfated GalNAc residue and two or three Man residues.     

Contributions of expression redistribution and allele substitutions to the genetic differentiation of animal taxa of different ranks

The contributions to the genetic differences between animal taxa of various ranks were estimated for four groups of loci: loci with identical (i), similar (s), or completely different (d) allele compositions, and (o) loci displaying a change in expression. The main contribution is made by i-and s-loci at the intraspecific level. Another pattern is observed at the specific level: d-loci become dominant and o-loci appear, suggesting fundamental changes in the genomes upon speciation. The relationships between the contributions of the four groups of loci are relatively stable within a class and differ among classes. Interestingly, the extent of genetic differentiation (as estimated from the proportions of the above four components) observed at the specific level in amphibians is achieved only at the level of families in birds. The approach is especially efficient for identifying hybrid populations and individuals.     

Orientierende Untersuchungen über die Einwirkung von gasförmigem Formaldehyd auf die grüne Pflanze

Ohne Zusammenfassung