Negative charges in the loop between the A and B box of the T1 domain are involved in Kv2.1 and Kv2.1/Kv6.3 channel tetramerization

C

The selectivity filter is involved in the U-type inactivation process of Kv2.1 and Kv3.1 channels

Voltage-gated potassium (Kv) channels display several types of inactivation processes, including N-, C-, and U-types. C-type inactivation is attributed to a nonconductive conformation of the selectivity filter (SF). It has been proposed that the activation gate and the channel's SF are allosterically coupled because the conformational changes of the former affect the structure of the latter and vice versa. The second threonine of the SF signature sequence (e.g., TTVGYG) has been proven to be essential for this allosteric coupling. To further study the role of the SF in U-type inactivation, we substituted the second threonine of the TTVGYG sequence by an alanine in the hKv2.1 and hKv3.1 channels, which are known to display U-type inactivation. Both hKv2.1-T377A and hKv3.1-T400A yielded channels that were resistant to inactivation, and as a result, they displayed noninactivating currents upon channel opening; i.e., hKv2.1-T377A and hKv3.1-T400A remained fully conductive upon prolonged moderate depolarizations, whereas in wild-type hKv2.1 and hKv3.1, the current amplitude typically reduces because of U-type inactivation. Interestingly, increasing the extracellular K+ concentration increased the macroscopic current amplitude of both hKv2.1-T377A and hKv3.1-T400A, which is similar to the response of the homologous T to A mutation in Shaker and hKv1.5 channels that display C-type inactivation. Our data support an important role for the second threonine of the SF signature sequence in the U-type inactivation gating of hKv2.1 and hKv3.1. SIGNIFICANCE Voltage-dependent K+ (Kv) channels generate cells' repolarizing power, which is consequently regulated by the channel's conductance. Aside from the opening or closure, Kv channels undergo inactivation that drives them into a lower or nonconductive state. Among the different inactivation processes described in Kv channels, the U-type process develops from a preopen but activated state. The molecular determinants of this process are, in contrast to the Ctype mechanism, not well characterized. Our data show that the intracellular part of the K+ selectivity filter within the pore domain is involved. An alanine for threonine substitution results in channels that do not inactivate upon opening, suggesting that an allosteric coupling between the activation gate and selectivity filter exists in U-type inactivation

WTC2005-63536 EFFECT OF CRYSTALLOGRAPHIC TEXTURE ON DEFORMATION FIELDS IN FRETTING CONTACTS

ABSTRACT Fretting contacts in the partial slip regime are simulated by a finite element model of a rigid cylinder on an elastic-crystal viscoplastic half-space. The half-space is modeled as duplex Ti-6Al-4V, a polycrystalline metal alloy consisting of equiaxed primary alpha grains and secondary lamellar alpha+beta grains. Various realistic 3-D crystallographic textures are considered. The deformation fields generated by fretting are quantified in terms of cumulative effective plastic strain distributions and plastic strain maps. The results clearly demonstrate the importance of the various sources of microstructural heterogeneity in the surface layers. The main sources of microstructural heterogeneity include the distribution of phases, slip system strength anisotropy, and crystallographic texture. In basal textured materials with fretting on the edge, the plastic strain is more evenly distributed in the subsurface regions than in other textured cases. This is explained by the greater number of grains able to deform by soft slip modes and the symmetry of this type of texture relative to the fretting orientation. Transverse and basal/transverse textures result in more heterogeneously-distributed plastic strain with strain often concentrated in narrow vein-like structures with maximum accumulation near alpha/alpha+beta grain boundaries. Elastic shakedown is more difficult to achieve in the later case. Ratcheting is the primary mechanism for cyclic plastic strain accumulation

WTC2005-63536 EFFECT OF CRYSTALLOGRAPHIC TEXTURE ON DEFORMATION FIELDS IN FRETTING CONTACTS

ABSTRACT Fretting contacts in the partial slip regime are simulated by a finite element model of a rigid cylinder on an elastic-crystal viscoplastic half-space. The half-space is modeled as duplex Ti-6Al-4V, a polycrystalline metal alloy consisting of equiaxed primary alpha grains and secondary lamellar alpha+beta grains. Various realistic 3-D crystallographic textures are considered. The deformation fields generated by fretting are quantified in terms of cumulative effective plastic strain distributions and plastic strain maps. The results clearly demonstrate the importance of the various sources of microstructural heterogeneity in the surface layers. The main sources of microstructural heterogeneity include the distribution of phases, slip system strength anisotropy, and crystallographic texture. In basal textured materials with fretting on the edge, the plastic strain is more evenly distributed in the subsurface regions than in other textured cases. This is explained by the greater number of grains able to deform by soft slip modes and the symmetry of this type of texture relative to the fretting orientation. Transverse and basal/transverse textures result in more heterogeneously-distributed plastic strain with strain often concentrated in narrow vein-like structures with maximum accumulation near alpha/alpha+beta grain boundaries. Elastic shakedown is more difficult to achieve in the later case. Ratcheting is the primary mechanism for cyclic plastic strain accumulation

Cats and Dogs, Hair and A Hero: A Quintet of New Milky Way Companions

We present five new satellites of the Milky Way discovered in Sloan Digital Sky Survey (SDSS) imaging data, four of which were followed-up with either the Subaru or the Isaac Newton Telescopes. They include four probable new dwarf galaxies -- one each in the constellations of Coma Berenices, Canes Venatici, Leo and Hercules -- together with one unusually extended globular cluster, Segue 1. We provide distances, absolute magnitudes, half-light radii and color-magnitude diagrams for all five satellites. The morphological features of the color-magnitude diagrams are generally well described by the ridge line of the old, metal-poor globular cluster M92. In the last two years, a total of ten new Milky Way satellites with effective surface brightness mu_v >~ 28 mag/sq. arcsec have been discovered in SDSS data. They are less luminous, more irregular and appear to be more metal-poor than the previously-known nine Milky Way dwarf spheroidals. The relationship between these objects and other populations is discussed. We note that there is a paucity of objects with half-light radii between ~40 pc and ~ 100 pc. We conjecture that this may represent the division between star clusters and dwarf galaxies.Comment: 10 pages, 8 figures, submitted to the Astrophysical Journa

The INT6 Cancer Gene and MEK Signaling Pathways Converge during Zebrafish Development

BACKGROUND: Int-6 (integration site 6) was identified as an oncogene in a screen of tumorigenic mouse mammary tumor virus (MMTV) insertions. INT6 expression is altered in human cancers, but the precise role of disrupted INT6 in tumorigenesis remains unclear, and an animal model to study Int-6 physiological function has been lacking. PRINCIPAL FINDINGS: Here, we create an in vivo model of Int6 function in zebrafish, and through genetic and chemical-genetic approaches implicate Int6 as a tissue-specific modulator of MEK-ERK signaling. We find that Int6 is required for normal expression of MEK1 protein in human cells, and for Erk signaling in zebrafish embryos. Loss of either Int6 or Mek signaling causes defects in craniofacial development, and Int6 and Erk-signaling have overlapping domains of tissue expression. SIGNIFICANCE: Our results provide new insight into the physiological role of vertebrate Int6, and have implications for the treatment of human tumors displaying altered INT6 expression