Regulation of Na+/K+ ATPase Transport Velocity by RNA Editing

Editing of Na+/K+ ATPase mRNAs modulates the Na+/K+ pump's turnover rate by selectively targeting the release of the final sodium to the outside

Refinement of 1p36 Alterations Not Involving PRDM16 in Myeloid and Lymphoid Malignancies

Fluorescence in situ hybridization was performed to characterize 81 cases of myeloid and lymphoid malignancies with cytogenetic 1p36 alterations not affecting the PRDM16 locus. In total, three subgroups were identified: balanced translocations (N = 27) and telomeric rearrangements (N = 15), both mainly observed in myeloid disorders; and unbalanced non-telomeric rearrangements (N = 39), mainly observed in lymphoid proliferations and frequently associated with a highly complex karyotype. The 1p36 rearrangement was isolated in 12 cases, mainly myeloid disorders. The breakpoints on 1p36 were more widely distributed than previously reported, but with identifiable rare breakpoint cluster regions, such as the TP73 locus. We also found novel partner loci on 1p36 for the known multi-partner genes HMGA2 and RUNX1. We precised the common terminal 1p36 deletion, which has been suggested to have an adverse prognosis, in B-cell lymphomas [follicular lymphomas and diffuse large B-cell lymphomas with t(14;18)(q32;q21) as well as follicular lymphomas without t(14;18)]. Intrachromosomal telomeric repetitive sequences were detected in at least half the cases of telomeric rearrangements. It is unclear how the latter rearrangements occurred and whether they represent oncogenic events or result from chromosomal instability during oncogenesis

Voltage-dependent charge movement associated with activation of the CLC-5 2Cl−/1H+ exchanger

The family of CLC proteins comprises both Cl− channels and Cl−/H+ exchange transporters with varying degrees of voltage dependence. The human CLC-5 is an electrogenic voltage-dependent 2Cl−/1H+ exchanger that gives rise to strongly outwardly rectifying currents when expressed. We conducted whole-cell recordings from HEK293 cells transiently transfected with either wild-type CLC-5 or a permeation-deficient mutant, E268A. With E268A CLC-5 we recorded transient voltage-dependent currents that represent the gating currents associated with CLC-5 activation and had kinetics that could be described by voltage-dependent forward and reverse transition rates. In extracellular solutions rich in Cl− or Br−, CLC-5 exhibited a gating charge of 1.3, but this was reduced to 0.9 in solutions comprising the impermeant anions aspartate, methanesulfonate, sulfate, or HEPES. Extracellular ion depletion by local perfusion with isotonic mannitol failed to reduce the gating charge further. Lowering intracellular pH from 7.4 to 5.4 did not shift the voltage-dependence of the gating currents, but reducing and increasing intracellular Cl− shifted the charge-voltage relationship to more negative and positive potentials, respectively. Our data suggest that voltage sensing is an intrinsic property of the CLC-5 protein and that permeant anions, particularly Cl−, modulate a voltage-dependent transition to an activated state from which Cl−/H+ exchange can occur.—Smith, A. J., Lippiat, J. D. Voltage-dependent charge movement associated with activation of the CLC-5 2Cl−/1H+ exchanger

Depolarization increases the apparent affinity of the Na+-K+ pump to cytoplasmic Na+ in isolated guinea-pig ventricular myocytes

In order to investigate the possible effect of membrane potential on cytoplasmic Na+ binding to the Na+-K+ pump, we studied Na+-K+ pump current-voltage relationships in single guinea-pig ventricular myocytes whole-cell voltage clamped with pipette solutions containing various concentrations of Na+ ([Na+]pip) and either tetraethylammonium (TEA+) or N-methyl-D-glucamine (NMDG+) as the main cation. The experiments were conducted at 30 °C under conditions designed to abolish the known voltage dependence of other steps in the pump cycle, i.e. in Na+-free external media containing 20 mM Cs+.Na+-K+ pump current (Ip) was absent in cells dialysed with Na+-free pipette solutions and was almost voltage independent at 50 mM Na+pip (potential range: −100 to +40 mV). By contrast, the activation of Ip by 0.5–5 mM Na+pip was clearly voltage sensitive and increased with depolarization, independently of the main intracellular cation species.The apparent affinity of the Na+-K+ pump for cytoplasmic Na+ increased monotonically with depolarization. The [Na+]pip required for half-maximal Ip activation (K0.5 value) amounted to 5.6 mM at −100 mV and to 2.2 mM at +40 mV.The results suggest that cytoplasmic Na+ binding and/or a subsequent partial reaction in the pump cycle prior to Na+ release is voltage dependent. From the voltage dependence of the K0.5 values the dielectric coefficient for intracellular Na+ binding/translocation was calculated to be ≈0.08. The voltage-dependent mechanism might add to the activation of the cardiac Na+-K+ pump during cardiac excitation