Cardiac expression of the cystic fibrosis transmembrane conductance regulator involves novel Exon 1 usage to produce a unique amino-terminal protein

Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which encodes a chloride channel present in many cells. In cardiomyocytes, we report that multiple exon 1 usage and alternative splicing produces four CFTR transcripts, with different 5'-untranslated regions, CFTRTRAD-139, CFTR-1C/-1A, CFTR-1C, and CFTR-1B. CFTR transcripts containing the novel upstream exons (exons -1C, -1B, and -1A) represent more than 90% of cardiac expressed CFTR mRNA. Regulation of cardiac CFTR expression, in response to developmental and pathological stimuli, is exclusively due to the modulation of CFTR-1C and CFTR-1C/-1A expression. Upstream open reading frames have been identified in the 5'-untranslated regions of all CFTR transcripts that, in conjunction with adjacent stem-loop structures, modulate the efficiency of translation initiation at the AUG codon of the main CFTR coding region in CFTRTRAD-139 and CFTR-1C/-1A transcripts. Exon(-1A), only present in CFTR-1C/-1A transcripts, encodes an AUG codon that is in-frame with the main CFTR open reading frame, the efficient translation of which produces a novel CFTR protein isoform with a curtailed amino terminus. As the expression of this CFTR transcript parallels the spatial and temporal distribution of the cAMP-activated whole-cell current density in normal and diseased hearts, we suggest that CFTR-1C/-1A provides the molecular basis for the cardiac cAMP-activated chloride channel. Our findings provide further insight into the complex nature of in vivo CFTR expression, to which multiple mRNA transcripts, protein isoforms, and post-transcriptional regulatory mechanisms are now added

Network adaptation improves temporal representation of naturalistic stimuli in drosophila eye: II Mechanisms

Retinal networks must adapt constantly to best present the ever changing visual world to the brain. Here we test the hypothesis that adaptation is a result of different mechanisms at several synaptic connections within the network. In a companion paper (Part I), we showed that adaptation in the photoreceptors (R1-R6) and large monopolar cells (LMC) of the Drosophila eye improves sensitivity to under-represented signals in seconds by enhancing both the amplitude and frequency distribution of LMCs' voltage responses to repeated naturalistic contrast series. In this paper, we show that such adaptation needs both the light-mediated conductance and feedback-mediated synaptic conductance. A faulty feedforward pathway in histamine receptor mutant flies speeds up the LMC output, mimicking extreme light adaptation. A faulty feedback pathway from L2 LMCs to photoreceptors slows down the LMC output, mimicking dark adaptation. These results underline the importance of network adaptation for efficient coding, and as a mechanism for selectively regulating the size and speed of signals in neurons. We suggest that concert action of many different mechanisms and neural connections are responsible for adaptation to visual stimuli. Further, our results demonstrate the need for detailed circuit reconstructions like that of the Drosophila lamina, to understand how networks process information

Evidence for a Prepore Stage in the Action of Clostridium perfringens Epsilon Toxin

Clostridium perfringens epsilon toxin (ETX) rapidly kills MDCK II cells at 37°C, but not 4°C. The current study shows that, in MDCK II cells, ETX binds and forms an oligomeric complex equally well at 37°C and 4°C but only forms a pore at 37°C. However, the complex formed in MDCK cells treated with ETX at 4°C has the potential to form an active pore, since shifting those cells to 37°C results in rapid cytotoxicity. Those results suggested that the block in pore formation at 4°C involves temperature-related trapping of ETX in a prepore intermediate on the MDCK II cell plasma membrane surface. Evidence supporting this hypothesis was obtained when the ETX complex in MDCK II cells was shown to be more susceptible to pronase degradation when formed at 4°C vs. 37°C; this result is consistent with ETX complex formed at 4°C remaining present in an exposed prepore on the membrane surface, while the ETX prepore complex formed at 37°C is unaccessible to pronase because it has inserted into the plasma membrane to form an active pore. In addition, the ETX complex rapidly dissociated from MDCK II cells at 4°C, but not 37°C; this result is consistent with the ETX complex being resistant to dissociation at 37°C because it has inserted into membranes, while the ETX prepore readily dissociates from cells at 4°C because it remains on the membrane surface. These results support the identification of a prepore stage in ETX action and suggest a revised model for ETX cytotoxicity, i) ETX binds to an unidentified receptor, ii) ETX oligomerizes into a prepore on the membrane surface, and iii) the prepore inserts into membranes, in a temperature-sensitive manner, to form an active pore

Measurement of D0 -> pilnu (Klnu) Form Factors and Absolute Branching Fractions

Using a 282 1/fb data sample collected by the Belle experiment at the KEKB e+e- collider, we study D0 decays to K-l+nu and pi-l+nu final states. The D0 flavor and momentum are tagged through a full reconstruction of the recoiling charm meson and additional mesons from fragmentation. The reconstruction method provides very good resolution in neutrino momentum and in q^2 = (p_l+p_nu)^2. Normalizing to the total number of D0 tags, we measure the absolute branching fractions to be B(D0 -> Klnu) =(3.45 +- 0.07stat +- 0.20syst)% and B(D0 -> pilnu) = (0.255 +- 0.019stat +- 0.016syst)% and the semi-leptonic form factors (within the modified pole model) f+^K(0) = 0.695 +- 0.007stat +- 0.022syst and f+^pi(0) = 0.624 +- 0.020stat +- 0.030syst.Comment: 9 pages, 2 figures, submitted to Phys. Rev. Let

Measurement of the branching fractions for B- --> D(*)+ pi- l- nu-bar and B0bar --> D(*)0 pi+ l- nu-bar

We report on a measurement of the branching fractions for B- --> D(*)+ pi- l- nu-bar and B0bar --> D(*)0 pi+ l- nu-bar with 275 million BBbar events collected at the Upsilon(4S) resonance with the Belle detector at KEKB. Events are tagged by fully reconstructing one of the B mesons in hadronic modes. We obtain Br(B- --> D+ pi- l- nu-bar) = (0.54 +/- 0.07(stat) +/- 0.07(syst) +/- 0.06(BR)) x 10^-2, Br(B- --> D*+ pi- l- nu-bar) = (0.67 +/- 0.11(stat) +/- 0.09(syst) +/- 0.03(BR)) x 10^-2, Br(B0bar --> D0 pi+ l- nu-bar) = (0.33 +/- 0.06(stat) +/- 0.06(syst) +/- 0.03(BR)) x 10^-2, Br(B0bar --> D*0 pi+ l- nu-bar) = (0.65 +/- 0.12(stat) +/- 0.08(syst) +/- 0.05(BR)) x 10^-2, where the third error comes from the error on Bbar --> D(*) l- nu-bar decays. Contributions from B0bar --> D*+ l- nu-bar decays are excluded in the measurement of B0bar --> D0 pi+ l- nu-bar.Comment: 6 pages, 10 figures, submitted to Physical Review D (Rapid Communication), the Lepton-Photon 2005 Conference (Uppsala, Sweden) and the HEP2005 Europhysics Conference (Lisboa, Portugal

Measurements of CP Violation in B0→D∗−π+B^0 \to D^{*-}\pi^+ and B0→D−π+B^0 \to D^- \pi^+ Decays

We report measurements of time dependent decay rates for $B^0 \to D^{(*)-}\pi^+$ decays and extraction of CP violation parameters that depend on $\phi_3$. Using fully reconstructed $D^{(*)}\pi$ events and partially reconstructed $D^{*}\pi$ events from a data sample that contains 386 million $B\bar{B}$ pairs that was collected near the $\Upsilon(4S)$ resonance, with the Belle detector at the KEKB asymmetric energy $e^+ e^-$ collider, we obtain the CP violation parameters $S^+ (D^{(*)}\pi)$ and $S^- (D^{(*)}\pi)$. We obtain $S^+ (D^* \pi) = 0.049 \pm 0.020(\mathrm{stat}) \pm 0.011(\mathrm{sys})$, $S^- (D^* \pi) = 0.031 \pm 0.019(\mathrm{stat}) \pm 0.011(\mathrm{sys})$, and $S^+ (D \pi) = 0.031 \pm 0.030(\mathrm{stat}) \pm 0.012(\mathrm{sys})$, $S^- (D \pi) = 0.068 \pm 0.029(\mathrm{stat}) \pm 0.012(\mathrm{sys})$. These results are an indication of CP violation in $B^0 \to D^{*-}\pi^+$ and $B^0 \to D^- \pi^+$ decays at the $2.5 \sigma$ and $2.2 \sigma$ levels, respectively. If we use the values of $R_{D^{(*)}\pi}$ that are derived using assumptions of factorization and SU(3) symmetry, the branching fraction measurements for the $D_s^{(*)} \pi$ modes, and lattice QCD calculations, we can restrict the allowed region of $|\sin (2\phi_1 + \phi_3)|$ to be above 0.44 and 0.52 at 68% confidence level from the $D^* \pi$ and $D \pi$ modes, respectively.Comment: 14 pages, 12 figures, submitted to Physical Review

Measurement of the wrong-sign decays D0 -> K+ pi- pi0 and D0 -> K+ pi- pi+ pi-, and search for CP violation

Using 281 fb^-1 of data from the Belle experiment recorded at or near the Upsilon(4S) resonance, we have measured the rates of the ``wrong-sign'' decays D0 -> K+ pi- pi0 and D0 -> K+ pi- pi+ pi- relative to those of the Cabibbo-favored decays D0 -> K- pi+ pi0 and D0 -> K- pi+ pi+ pi-. These wrong-sign decays proceed via a doubly Cabibbo-suppressed amplitude or via D0-D0bar mixing; the latter has not yet been observed. We obtain R_WS(Kpipi0)=[0.229 +/-0.017(stat.) +0.013-0.009(sys.)]% and R_WS(K3pi)=[0.320 +/-0.019(stat.) +0.018-0.013(sys.)]%. The CP asymmetries are measured to be -0.006 +/- 0.053 and -0.018 +/- 0.044 for the K+ pi- pi0 and K+ pi- pi+ pi- final states, respectively.Comment: 10 pages, 3 figures, submitted to PRL, Lepton-Photon 2005 Conference in Uppsala, Sweden and HEP2005 Europhysics Conference in Lisboa, Portuga

Measurement of phi_3 with Dalitz plot analysis of B+ -> D(*)K(*)+ decay

We present a measurement of the unitarity triangle angle phi_3 using a Dalitz plot analysis of the K0_S pi+ pi- decay of the neutral D meson from the B+- -> D(*)K(*)+- process. The method employs the interference between D0 and D0bar to extract the angle phi_3, strong phase Delta and the ratio r of suppressed and allowed amplitudes. We apply this method to a 357 fb-1 data sample collected by the Belle experiment. The analysis uses three modes: B+ -> DK+, B+ -> D*K+ with D* -> Dpi0, and B+ -> DK*+ with K*+ -> K0_S pi+, as well as the corresponding charge-conjugate modes. From a combined maximum likelihood fit to the three modes, we obtain phi_3=53+15-18(stat)+-3(syst)+-9(model) degrees. The corresponding two standard deviation interval is 8<phi_3<111 degrees.Comment: 18 pages, 6 figures, 7 tables. To be submitted to Phys. Rev.

Genetic plasticity of the Shigella virulence plasmid is mediated by intra- and inter-molecular events between insertion sequences

Acquisition of a single copy, large virulence plasmid, pINV, led to the emergence of Shigella spp. from Escherichia coli. The plasmid encodes a Type III secretion system (T3SS) on a 30kb pathogenicity island (PAI), and is maintained in a bacterial population through a series of toxin:antitoxin (TA) systems which mediate post-segrega tional killing (PSK). The T3SS imposes a significant cost on the bacterium, and strains which have lost the plasmid and/or genes encoding the T3SS grow faster than wild-type strains in the laboratory, and fail to bind the indicator dye Congo Red (CR). Our aim was to define the molecular events in Shigella flexneri that cause loss of Type III secretion (T3S), and to examine whether TA systems exert positional effects on pINV. During growth at 37°C, we found that deletions of regions of the plasmid including the PAI lead to the emergence of CR-negative colonies; deletions occur through intra-molecula r recombination events between insertion sequences (ISs) flanking the PAI. Furthermore, by repositioning MvpAT (which belongs to the VapBC family of TA systems) near the PAI, we demonstrate that the location of this TA system alters the rearrangements that lead to loss of T3S, indicating that MvpAT acts both globally (by reducing loss of pINV through PSK) as well as locally (by preventing loss of adjacent sequences). During growth at environmental temperatures, we show for the first time that pINV spontaneously integrates into different sites in the chromosome, and this is mediated by inter-molecular events involving IS 1294. Integration leads to reduced PAI gene expression and impaired secretion through the T3SS, while excision of pINV from the chromosome restores T3SS function. Therefore, pINV integration provides a reversible mechanism for Shigella to circumvent the metabolic burden imposed by pINV. Intra- and inter-molecular events between ISs, which are abundant in Shigella spp., mediate plasticity of S. flexneri pINV