Dual lipid modification of the yeast ggamma subunit Ste18p determines membrane localization of Gbetagamma

The pheromone response in the yeast Saccharomyces cerevisiae is mediated by a heterotrimeric G protein. The Gbetagamma subunit (a complex of Ste4p and Ste18p) is associated with both internal and plasma membranes, and a portion is not stably associated with either membrane fraction. Like Ras, Ste18p contains a farnesyl-directing CaaX box motif (C-terminal residues 107 to 110) and a cysteine residue (Cys 106) that is a potential site for palmitoylation. Mutant Ste18p containing serine at position 106 (mutation ste18-C106S) migrated more rapidly than wild-type Ste18p during sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The electrophoretic mobility of wild-type Ste18p (but not the mutant Ste18p) was sensitive to hydroxylamine treatment, consistent with palmitoyl modification at Cys 106. Furthermore, immunoprecipitation of the Gbetagamma complex from cells cultured in the presence of [(3)H]palmitic acid resulted in two radioactive species on nonreducing SDS-PAGE gels, with molecular weights corresponding to Ggamma and Gbetagamma. Substitution of serine for either Cys 107 or Cys 106 resulted in the failure of Gbetagamma to associate with membranes. The Cys 107 substitution also resulted in reduced steady-state accumulation of Ste18p, suggesting that the stability of Ste18p requires modification at Cys 107. All of the mutant forms of Ste18p formed complexes with Ste4p, as assessed by coimmunoprecipitation. We conclude that tight membrane attachment of the wild-type Gbetagamma depends on palmitoylation at Cys 106 and prenylation at Cys 107 of Ste18p

Interaction Among \u3cem\u3eSaccharomyces cerevisiae\u3c/em\u3e Pheromone Receptors During Endocytosis

This study investigates endocytosis of Saccharomyces cerevisiae α-factor receptor and the role that receptor oligomerization plays in this process. α-factor receptor contains signal sequences in the cytoplasmic C-terminal domain that are essential for ligand-mediated endocytosis. In an endocytosis complementation assay, we found that oligomeric complexes of the receptor undergo ligand-mediated endocytosis when the α-factor binding site and the endocytosis signal sequences are located in different receptors. Both in vitro and in vivo assays suggested that ligand-induced conformational changes in one Ste2 subunit do not affect neighboring subunits. Therefore, recognition of the endocytosis signal sequence and recognition of the ligand-induced conformational change are likely to be two independent events

Feasibility study of the transonic biplane concept for transport aircraft application

Investigations were conducted to evaluate the feasibility of a transonic biplane consisting of a forward-mounted swept-back lower wing, a rear-mounted swept-forward upper wing, and a vertical fin connecting the wings at their tips. This wing arrangement results in significant reductions in induced drag relative to a monoplane designed with the same span, and it allows for a constant-section fuselage shape while closely matching an ideal area distribution curve for M = 0.95 cruise. However, no significant reductions in ramp weight were achieved for the biplane relative to a monoplane with the same mission capability. Flutter analyses of the biplane revealed both symmetric and antisymmetric instabilities that occur well below the required flutter speed. Further studies will be required to determine if acceptable flutter speeds can be achieved through the elimination of the instabilities by passive means or by active controls. Configurations designed for other missions, especially those with lower Mach numbers and lower dynamic pressures, should be examined since the geometries suitable for those design constraints might avoid the weight penalties and flutter instabilities which prevent exploitation of induced drag benefits for the configuration studied

The relationship of parental discipline style to gender differences in early adolescent compliance, self-image and peer relations :: a comparison of child and parent perspectives.

Thesis (M.S.

Resolving the Submillimeter Background: the 850-micron Galaxy Counts

Recent deep blank field submillimeter surveys have revealed a population of luminous high redshift galaxies that emit most of their energy in the submillimeter. The results suggest that much of the star formation at high redshift may be hidden to optical observations. In this paper we present wide-area 850-micron SCUBA data on the Hawaii Survey Fields SSA13, SSA17, and SSA22. Combining these new data with our previous deep field data, we establish the 850-micron galaxy counts from 2 mJy to 10 mJy with a >3-sigma detection limit. The area coverage is 104 square arcmin to 8 mJy and 7.7 square arcmin to 2.3 mJy. The differential 850-micron counts are well described by the function n(S)=N_0/(a+S^3.2), where S is the flux in mJy, N_0=3.0 x 10^4 per square degree per mJy, and a=0.4-1.0 is chosen to match the 850-micron extragalactic background light. Between 20 to 30 per cent of the 850-micron background resides in sources brighter than 2 mJy. Using an empirical fit to our >2 mJy data constrained by the EBL at lower fluxes, we argue that the bulk of the 850-micron extragalactic background light resides in sources with fluxes near 1 mJy. The submillimeter sources are plausible progenitors of the present-day spheroidal population.Comment: 5 pages, accepted by The Astrophysical Journal Letter

CHIMPS: the ¹³CO/C¹⁸O (<i>J</i> = 3 → 2) Heterodyne Inner Milky Way Plane Survey

We present the 13CO/C18O (J = 3 → 2) Heterodyne Inner Milky Way Plane Survey (CHIMPS) which has been carried out using the Heterodyne Array Receiver Program on the 15 m James Clerk Maxwell Telescope (JCMT) in Hawaii. The high-resolution spectral survey currently covers |b| ≤ 0.5° and 28° ≲ l ≲ 46°, with an angular resolution of 15 arcsec in 0.5 km s-1 velocity channels. The spectra have a median rms of ˜0.6 K at this resolution, and for optically thin gas at an excitation temperature of 10 K, this sensitivity corresponds to column densities of NH2 ˜ 3 × 1020 cm-2 and NH2 ˜ 4 × 1021 cm-2 for 13CO and C18O, respectively. The molecular gas that CHIMPS traces is at higher column densities and is also more optically thin than in other publicly available CO surveys due to its rarer isotopologues, and thus more representative of the three-dimensional structure of the clouds. The critical density of the J = 3 → 2 transition of CO is ≳104 cm-3 at temperatures of ≤20 K, and so the higher density gas associated with star formation is well traced. These data complement other existing Galactic plane surveys, especially the JCMT Galactic Plane Survey which has similar spatial resolution and column density sensitivity, and the Herschel infrared Galactic Plane Survey. In this paper, we discuss the observations, data reduction and characteristics of the survey, presenting integrated-emission maps for the region covered. Position-velocity diagrams allow comparison with Galactic structure models of the Milky Way, and while we find good agreement with a particular four-arm model, there are some significant deviations

Increased accuracy of ligand sensing by receptor internalization

Many types of cells can sense external ligand concentrations with cell-surface receptors at extremely high accuracy. Interestingly, ligand-bound receptors are often internalized, a process also known as receptor-mediated endocytosis. While internalization is involved in a vast number of important functions for the life of a cell, it was recently also suggested to increase the accuracy of sensing ligand as the overcounting of the same ligand molecules is reduced. Here we show, by extending simple ligand-receptor models to out-of-equilibrium thermodynamics, that internalization increases the accuracy with which cells can measure ligand concentrations in the external environment. Comparison with experimental rates of real receptors demonstrates that our model has indeed biological significance.Comment: 9 pages, 4 figures, accepted for publication in Physical Review

To Contact Tracing... And Beyond!

To the Editor—We write to endorse and extend the thoughts of Nosyk et al on contact tracing, which we strongly agree is an opportunity to achieve additional public health goals, including diagnosis, treatment, and prevention of human immunodeficiency virus (HIV)

The JCMT Gould Belt survey: Dense core clusters in Orion B

The James Clerk Maxwell Telescope Gould Belt Legacy Survey obtained SCUBA-2 observations of dense cores within three sub-regions of OrionB: LDN1622, NGC2023/2024, and NGC2068/2071, all of which contain clusters of cores. We present an analysis of the clustering properties of these cores, including the two-point correlation function and Cartwright’s Q parameter. We identify individual clusters of dense cores across all three regions using a minimal spanning tree technique, and find that in each cluster, the most massive cores tend to be centrally located. We also apply the independent M–Σ technique and find a strong correlation between core mass and the local surface density of cores. These two lines of evidence jointly suggest that some amount of mass segregation in clusters has happened already at the dense core stage