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Structural implications of the EL(K/Q)(L/C)LD(A/G)DD sequence in the C-terminal cytoplasmic tail for proper targeting of anion exchanger 1 to the plasma membrane
2009
Adachi, H., Hokkaido Univ., Sapporo (Japan) | Ito, D. | Kurooka, T. | Otsuka, Y. | Arashiki, N. | Sato, K. | Inaba, M.
While the C-terminal cytoplasmic tail of anion exchanger 1 (AE1, band 3) has been reported to possess important physiological roles, including one for proper membrane trafficking, its precise characteristics remain unclear. To clarify the overall structural consequences of the conserved sequence EL(K/Q)(L/C)LD(A/G)DD, containing the core binding sequence LDADD for carbonic anhydrase II, in the C-terminal region, we analyzed the membrane expression and turnover of bovine AE1 with a series of truncation and substitution mutations in HEK293 cells. Immunofluorescence microscopy and cell-surface biotinylation demonstrated that truncation mutants missing 18 C-terminal residues targeted the plasma membrane, but the one lacking the conserved region, by truncation of 28 amino acid residues, was retained inside the cells. Substitutions of Ala for Glusup(901), Leusup(902), Leusup(905), and Aspsup(906) in the sequence E901L(K/Q)(L/C)LDADD909 of bovine AE1 or those in the corresponding murine sequence also caused intracellular retention, though these mutants had half-lives comparable to that for wild-type AE1. These data demonstrate that the conserved amino acid residues Glusup(1), Leusup(2), Leusup(5), and Aspsup(6) in the EL(K/Q)(L/C)LD(A/G)DD region have essential structural consequences in stable expression of AE1 at the plasma membrane regardless of the ability in binding to carbonic anhydrase II of this region.
Показать больше [+] Меньше [-]Radioecological investigation of food of animal origin in Belgrade environment
2009
Gordana, V., Belgrade Univ. (Serbia) | Svetlana, G. | Branislava, M. | Milan, O. | Branko, P.
The activity concentrations of sup(40)K, sup(238)U, sup(232)Th and sup(137)Cs were measured using gamma spectrometric method in different food chain samples from Serb Belgrade environment during the periods May-June 2007 and May-June 2008 year. Relatively high activities of sup(40)K and sup(137)Cs were detected in the soil. These results indicate that sup(137)Cs is present in Belgrade environment even 20 years after nuclear accident in Chernobyl. However, in the samples of feedstuffs, animal products and bio indicators (meat of wild animals and fish), activity concentrations of primordial radionuclides and sup(137)Cs were low and below the detection limits. Results of these trials have shown that investigated animal products from the natural environment around Belgrade, are radioactivity safe.
Показать больше [+] Меньше [-]In vitro propagation of rabies virus in mouse dorsal root ganglia cells
2009
Hara, Y.(Hokkaido Univ., Sapporo (Japan)) | Sunden, Y. | Ochiai, K. | Umemura, T.
Rabies virus (RV) is highly neurotropic and migrates to the neuronal soma by retrograde axonal transport from nerve terminals, after which it is taken by anterograde axonal transport to be finally released into the central nervous system (CNS) from which it disseminates, resulting in lethal encephalitis. Dorsal root ganglia (DRG) are crucial in the initial events of the infection by RV since they can act as a gate for the viral entrance into the CNS. In the present study, we examined cell tropism of RV and the roles of neuronal cytoskeletal components in the production of viral nucleoprotein (N protein) using cultured nerve cells and non-neuronal cells from DRG of newborn mice. Our in vitro study demonstrated a low propagation rate of RV in nerve cells, susceptibility of non-neuronal cells to RV, and independence of cytoplasmic synthesis of viral N protein from the neuronal cytoskeleton. The present study also suggests that Schwann cells should be considered as another possible candidate supporting RV propagation.
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