The positive-acting sulfur regulatory protein CYS3 of Neurospora crassa: nuclear localization, autogenous control, and regions required for transcriptional activation

The positive-acting global sulfur regulatory protein, CYS3, of Neurospora crassa turns on the expression of a family of unlinked structural genes that encode enzymes of sulfur catabolism. CYS3 contains a leucine zipper and an adjacent basic region (b-zip), which together constitute a bipartite sequence-specific DNA-binding domain. Specific anti-CYS3 antibodies detected a protein of the expected size in nuclear extracts of wild-type Neurospora under conditions in which the sulfur circuit is activated. The CYS3 protein was not observed in cys-3 mutants. Nuclear extracts of wild type, but not cys-3 mutants, also showed specific DNA-binding activity identical to that obtained with a CYS3 protein expressed in Escherichia coli. A truncated CYS3 protein that contains primarily the b-zip domain binds to DNA with high specificity and affinity in vitro, yet fails to activate gene expression in vivo, and instead inhibits the function of the wild-type CYS3 protein. Amino-terminal, carboxyterminal, and internal deletions as well as alanine scanning mutagenesis were employed to identify regions of the CYS3 protein that are required for its trans-activation function. Regions of CYS3 carboxy terminal to the b-zip motif are not completely essential for function although loss of an alanine-rich region results in decreased activity. All deletions amino terminal to the b-zip motif led to a complete loss of CYS3 function. Alanine scanning mutagenesis demonstrated that an unusual prolinerich domain of CYS3 appears to be very important for function and is presumed to constitute an activation domain. It is concluded that CYS3 displays nuclear localization and positive autogenous control in Neurospora and functions as a trans-acting DNA-binding protein.     

Lobectomy combined with volume reduction for patients with lung cancer and advanced emphysema

Methanol-induced conformational transitions of hen egg white lysozyme were investigated with a combined use of far- and near-UV CD and NMR spectroscopies, ANS binding and small-angle X-ray scattering. Addition of methanol induced no global change in the native conformation itself, but induced a transition from the native state to the denatured state which was highly cooperative, as shown by the coincidence of transition curves monitored by the far- and near-UV CD spectroscopy, by isodichroic points in the far- and near-UV CD spectra and by the concomitant disappearance of individual 1H NMR signals of the native state. The ANS binding experiments could detect no intermediate conformer similar to the molten globule state in the process of the methanol denaturation. However, at high concentration of methanol, e.g., 60% (v/v) methanol/water, a highly helical state (H) was realized. The H state had a helical content much higher than the native state, monitored by far-UV CD spectroscopy, and had no specific tertiary structure, monitored both by near-UV CD and NMR spectroscopy. The radius of gyration in the H state, 24.9 angstroms, was significantly larger than that in the native state (15.7 angstroms). The Kratky plot for the H state did not show a clear peak and was quite similar to that for the urea-denatured state, indicating a complete lack of globularity. Thus we conclude that the H state has a considerably expanded, flexible broken rod-like conformation which is clearly distinguishable from the "molten globule" state. The stability of both N and H states depends on pH and methanol concentration. Thus a phase diagram involving N and H was constructed.     

Characterization of natriuretic activity from posterior pituitary lobes

Previous studies have indicated the existence of natriuretic factors of hormonal nature with the posterior pituitary gland as a possible site of origin. It was in this light that a series of experiments was designed to examine the posterior pituitary for such factors. Acetic acid extracts of porcine and bovine posterior pituitary lobe tissue were subjected to gel filtration on Sephadex G-25. Several fractions in the molecular size range of 1000 were obtained which possessed potent natriuretic activity as assayed in rats. The activity of these fractions maximally increased sodium excretion to 6-8 muequiv./min, a 10- to 40-fold increase above control, when administered intraperitoneally to hydropenic, conscious rats. However, oxytocin and vasopressin, present in the posterior pituitary are natriuretic. These hormones were measured by radioimmunoassay, and invariably only those fractions which contained vasopressin and (or) oxytocin possessed natriuretic activity. Moreover, the extent of the natriuresis could be accounted for by the vasopressin and (or) oxytocin content of the test fractions. The natriuretic property of this material was abolished by treatment with thioglycollate. Further purification of natriuretic fractions by ion exchange resins, thin-layer chromatography and isoelectric focusing failed to resolve natriuretic activity from vasopressin and oxytocin. Similar results were observed following analysis of fractions isolated by gel filtration of acetic acid extracts of ventral hypothalamus tissue. The natriuretic fractions isolated from hypothalamic tissue were indistinguishable from oxytocin and vasopressin. These experiments suggest that the natriuretic activity in neurohypophyseal extracts can be attributed to oxytocin and vasopressin.     

Associations between polymorphic variety and anthropometric and biochemical variation in two New Guinea populations

During International Biological Programme studies in Papua New Guinea, on Karkar Island and at Lufa in the Eastern Highlands, information on anthropometric, biochemical and genetic characteristics was collected from the same individuals. Use has been made of this special situation to investigate whether any associations exist between genetic markers and anthropometric and biochemical variation. Those found, and which satisfy criteria established to help in distinguishing real from chance associations, include: P with haemoglobin concentration and serum albumin and cholesterol levels; acid phosphatase with serum albumin level and haemoglobin concentration; anhaptoglobinaemia and serum globulin level; haemoglobin J-Tongariki with serum phosphorus level. The associations are discussed in terms of their arising from the direct result of environmental factors modifying gene expression, chance concordance of environmental and genetic variation, or pleiotropy.     

Digital low-pass filtering

Random noise beyond the maximum signal frequency of the imaging system can be removed by low-pass filtering without compromising signal transmission.