Genetic and biochemical analysis of the adenylyl cyclase-associated protein, cap, in Schizosaccharomyces pombe

We have identified, cloned, and studied a gene, cap, encoding a protein that is associated with adenylyl cyclase in the fission yeast Schizosaccharomyces pombe. This protein shares significant sequence homology with the adenylyl cyclase-associated CAP protein in the yeast Saccharomyces cerevisiae. CAP is a bifunctional protein; the N-terminal domain appears to be involved in cellular responsiveness to RAS, whereas loss of the C-terminal portion is associated with morphological and nutritional defects. S. pombe cap can suppress phenotypes associated with deletion of the C-terminal CAP domain in S. cerevisiae but does not suppress phenotypes associated with deletion of the N-terminal domain. Analysis of cap disruptants also mapped the function of cap to two domains. The functional loss of the C-terminal region of S. pombe cap results in abnormal cellular morphology, slow growth, and failure to grow at 37-degrees-C. Increases in mating and sporulation were observed when the entire gene was disrupted. Overproduction of both cap and adenylyl cyclase results in highly elongated large cells that are sterile and have measurably higher levels of adenylyl cyclase activity. Our results indicate that cap is required for the proper function of S. pombe adenylyl cyclase but that the C-terminal domain of cap has other functions that are shared with the C-terminal domain of S. cerevisiae CAP

Automation of Pivot Sprinkler Irrigation Systems to More Efficiently Utilize Rainfall and Irrigation Water

A study was conducted to develop automated pivot sprinkler irrigation systems and determine if such systems use less water and energy than manually operated systems. The study was conducted near Earth, Texas, using irrigation systems located on producers farms. Sensors with transmitters and receivers were constructed and tested so that the irrigation systems can be controlled by wind, soil water tension, and rainfall. The sensors can be used separately or in combination to control the irrigation systems. For several reasons it was not possible to determine if automated systems use less water and energy than manually operated systems. The major reason was the low capacity of the wells (114 to 204 m3/hr) supplying the irrigation systems. To meet crop water requirements and losses due to evaporation and runoff, the well capacity should be at least 284 m3/hr. Since the wells could not supply adequate water, soil water tension was out of the tensiometer range for the last 60 days of the growing season. Considerable variation in soil water tension and content was noted between irrigation systems and within quadrants of each irrigation system. Systems planted to cotton would probably be easier to automate than those planted in corn because of the lower water requirements of cotton. The wind and rainfall controls have more promise to aid in increasing water use efficiency than controls activated by soil water sensors. Wind controls could be used during preirrigation when more time is available to apply water and rainfall controls could be an aid to producers with remotely located irrigation systems

Negative regulation of syntaxin4/SNAP-23/VAMP2-mediated membrane fusion by Munc18c <i>In Vitro</i>

Background: Translocation of the facilitative glucose transporter GLUT4 from an intracellular store to the plasma membrane is responsible for the increased rate of glucose transport into fat and muscle cells in response to insulin. This represents a specialised form of regulated membrane trafficking. Intracellular membrane traffic is subject to multiple levels of regulation by conserved families of proteins in all eukaryotic cells. Notably, all intracellular fusion events require SNARE proteins and Sec1p/Munc18 family members. Fusion of GLUT4-containing vesicles with the plasma membrane of insulin-sensitive cells involves the SM protein Munc18c, and is regulated by the formation of syntaxin 4/SNAP23/VAMP2 SNARE complexes. Methodology/Principal Findings Here we have used biochemical approaches to characterise the interaction(s) of Munc18c with its cognate SNARE proteins and to examine the role of Munc18c in regulating liposome fusion catalysed by syntaxin 4/SNAP23/VAMP2 SNARE complex formation. We demonstrate that Munc18c makes contacts with both t- and v-SNARE proteins of this complex, and directly inhibits bilayer fusion mediated by the syntaxin 4/SNAP23/VAMP2 SNARE complex. Conclusion/Significance Our reductionist approach has enabled us to ascertain a direct inhibitory role for Munc18c in regulating membrane fusion mediated by syntaxin 4/SNAP23/VAMP2 SNARE complex formation. It is important to note that two different SM proteins have recently been shown to stimulate liposome fusion mediated by their cognate SNARE complexes. Given the structural similarities between SM proteins, it seems unlikely that different members of this family perform opposing regulatory functions. Hence, our findings indicate that Munc18c requires a further level of regulation in order to stimulate SNARE-mediated membrane fusion

Hearing shapes of drums - mathematical and physical aspects of isospectrality

In a celebrated paper '"Can one hear the shape of a drum?"' M. Kac [Amer. Math. Monthly 73, 1 (1966)] asked his famous question about the existence of nonisometric billiards having the same spectrum of the Laplacian. This question was eventually answered positively in 1992 by the construction of noncongruent planar isospectral pairs. This review highlights mathematical and physical aspects of isospectrality.Comment: 42 pages, 60 figure

Shell evolution of N = 40 isotones towards 60Ca: First spectroscopy of 62Ti

Excited states in the N=40 isotone 62Ti were populated via the 63V(p,2p)62Ti reaction at ∼200 MeV/nucleon at the Radioactive Isotope Beam Factory and studied using γ-ray spectroscopy. The energies of the 21+→0gs+ and 41+→21+ transitions, observed here for the first time, indicate a deformed 62Ti ground state. These energies are increased compared to the neighboring 64Cr and 66Fe isotones, suggesting a small decrease of quadrupole collectivity. The present measurement is well reproduced by large-scale shell-model calculations based on effective interactions, while ab initio and beyond mean-field calculations do not yet reproduce our findings. The shell-model calculations for 62Ti show a dominant configuration with four neutrons excited across the N=40 gap. Likewise, they indicate that the N=40 island of inversion extends down to Z=20, disfavoring a possible doubly magic character of the elusive 60Ca

Octupole states in Tl-207 studied through beta decay

The beta decay of Hg-207 into the single-proton-hole nucleus Tl-207 has been studied through gamma-ray spectroscopy at the ISOLDE Decay Station (IDS) with the aim of identifying states resulting from coupling of the pi s(1/2)(-1), pi d(3/2)(-1) and pi h(11/2)(-1) shell model orbitals to the collective octupole vibration. Twenty-two states were observed lying between 2.6 and 4.0 MeV, eleven of which were observed for the first time, and 78 new transitions were placed. Two octupole states (s(3/2)-coupled) are identified and three more states (d(3/2)-coupled) are tentatively assigned using spin-parity inferences, while further h(11/2)-coupled states may also have been observed for the first time. Comparisons are made with state-of-the-art large-scale shell model calculations and previous observations made in this region, and systematic underestimation of the energy of the octupole vibrational states is noted. We suggest that in order to resolve the difference in predicted energies for collective and noncollective t = 1 states (t is the number of nucleons breaking the Pb-208 core), the effect of t = 2 mixing may be reduced for octupole-coupled states. The inclusion of mixing with t = 0, 2, 3 excitations is necessary to replicate all t = 1 state energies accurately.Peer reviewe

Competition between Allowed and First-Forbidden beta Decay : The Case of Hg-208 -> Tl-2(0)8

The beta decay of Hg-208 into the one-proton hole, one neutron-particle Tl-208(81)127 nucleus was investigated at CERN-ISOLDE. Shell-model calculations describe well the level scheme deduced, validating the proton-neutron interactions used, with implications for the whole of the N > 126, Z 0(-)beta decay where the daughter state is core excited is unique, and can provide information on mesonic corrections of effective operators.Peer reviewe

Stab Injury to the Preauricular Region With Laceration of the External Carotid Artery Without Involvement of the Facial Nerve: a Case Report

BACKGROUND: Open injuries to the face involving the external carotid artery are uncommon. These injuries are normally associated with laceration of the facial nerve because this nerve is more superficial than the external carotid artery. Hence, external carotid artery lesions are usually associated with facial nerve dysfunction. We present an unusual case report in which the patient had an injury to this artery with no facial nerve compromise. CASE PRESENTATION: A 25-year-old Portuguese man sustained a stab wound injury to his right preauricular region with a broken glass. Immediate profuse bleeding ensued. Provisory tamponade of the wound was achieved at the place of aggression by two off-duty doctors. He was initially transferred to a district hospital, where a large arterial bleeding was observed and a temporary compressive dressing was applied. Subsequently, the patient was transferred to a tertiary hospital. At admission in the emergency room, he presented a pulsating lesion in the right preauricular region and slight weakness in the territory of the inferior buccal branch of the facial nerve. The physical examination suggested an arterial lesion superficial to the facial nerve. However, in the operating theater, a section of the posterior and lateral flanks of the external carotid artery inside the parotid gland was identified. No lesion of the facial nerve was observed, and the external carotid artery was repaired. To better understand the anatomical rationale of this uncommon clinical case, we dissected the preauricular region of six cadavers previously injected with colored latex solutions in the vascular system. A small triangular space between the two main branches of division of the facial nerve in which the external carotid artery was not covered by the facial nerve was observed bilaterally in all cases. CONCLUSIONS: This clinical case illustrates that, in a preauricular wound, the external carotid artery can be injured without facial nerve damage. However, no similar description was found in the reviewed literature, which suggests that this must be a very rare occurrence. According to the dissection study performed, this is due to the existence of a triangular space between the cervicofacial and temporofacial nerve trunks in which the external carotid artery is not covered by the facial nerve or its branches.info:eu-repo/semantics/publishedVersio