Local actin nucleation tunes centrosomal microtubule nucleation during passage through mitosis

Cells going through mitosis undergo precisely timed changes in cell shape and organisation, which serve to ensure the fair partitioning of cellular components into the two daughter cells. These structural changes are driven by changes in actin filament and microtubule dynamics and organisation. While most evidence suggests that the two cytoskeletal systems are remodelled in parallel during mitosis, recent work in interphase cells has implicated the centrosome in both microtubule and actin nucleation, suggesting the potential for regulatory crosstalk between the two systems. Here, by using both in vitro and in vivo assays to study centrosomal actin nucleation as cells pass through mitosis, we show that mitotic exit is accompanied by a burst in cytoplasmic actin filament formation that depends on WASH and the Arp2/3 complex. This leads to the accumulation of actin around centrosomes as cells enter anaphase and to a corresponding reduction in the density of centrosomal microtubules. Taken together, these data suggest that the mitotic regulation of centrosomal WASH and the Arp2/3 complex controls local actin nucleation, which may function to tune the levels of centrosomal microtubules during passage through mitosis

Monoubiquitination of syntaxin 3 leads to retrieval from the basolateral plasma membrane and facilitates cargo recruitment to exosomes

Syntaxin 3 (Stx3), a SNARE protein located and functioning at the apical plasma membrane of epithelial cells, is required for epithelial polarity. A fraction of Stx3 is localized to late endosomes/lysosomes, although how it traffics there and its function in these organelles is unknown. Here we report that Stx3 undergoes monoubiquitination in a conserved polybasic domain. Stx3 present at the basolateral—but not the apical—plasma membrane is rapidly endocytosed, targeted to endosomes, internalized into intraluminal vesicles (ILVs), and excreted in exosomes. A nonubiquitinatable mutant of Stx3 (Stx3-5R) fails to enter this pathway and leads to the inability of the apical exosomal cargo protein GPRC5B to enter the ILV/exosomal pathway. This suggests that ubiquitination of Stx3 leads to removal from the basolateral membrane to achieve apical polarity, that Stx3 plays a role in the recruitment of cargo to exosomes, and that the Stx3-5R mutant acts as a dominant-negative inhibitor. Human cytomegalovirus (HCMV) acquires its membrane in an intracellular compartment and we show that Stx3-5R strongly reduces the number of excreted infectious viral particles. Altogether these results suggest that Stx3 functions in the transport of specific proteins to apical exosomes and that HCMV exploits this pathway for virion excretion

PARP inhibitors as P-glyoprotein substrates

The cytotoxicity of PARP inhibitors olaparib, veliparib, and CEP-8983 were investigated in two P-glycoprotein (P-gp) overexpressing drug-resistant cell models (IGROVCDDP and KB-8-5-11). IGROVCDDP and KB-8-5-11 were both resistant to olaparib and resistance was reversible with the P-gp inhibitors elacridar, zosuquidar, and valspodar. In contrast, the P-gp overexpressing models were not resistant to veliparib or CEP-8983. Olaparib and veliparib did not induce protein expression of P-gp in IGROVCDDP or KB-8-5-11 at doses that successfully inhibit PARP. Olaparib therefore appears to be a P-gp substrate. Veliparib and CEP-8983 do not appear to be substrates. Veliparib and CEP-8983 may therefore be more useful in combined chemotherapy regimens with P-gp substrates and may be active in platinum and taxane-resistant ovarian cancer

New mechanism for Notch signaling to endothelium at a distance by Delta-like 4 incorporation into exosomes.

Notch signaling is an evolutionary conserved pathway that is mediated by cell-cell contact. It is involved in a variety of developmental processes and has an essential role in vascular development and angiogenesis. Delta-like 4 (Dll4) is a Notch ligand that is up-regulated during angiogenesis. It is expressed in endothelial cells and regulates the differentiation between tip cells and stalk cells of neovasculature. Here, we present evidence that Dll4 is incorporated into endothelial exosomes. It can also be incorporated into the exosomes of tumor cells that overexpress Dll4. These exosomes can transfer the Dll4 protein to other endothelial cells and incorporate it into their cell membrane, which results in an inhibition of Notch signaling and a loss of Notch receptor. Transfer of Dll4 was also shown in vivo from tumor cells to host endothelium. Addition of Dll4 exosomes confers a tip cell phenotype on the endothelial cell, which results in a high Dll4/Notch-receptor ratio, low Notch signaling, and filopodia formation. This was further evidenced by increased branching in a tube-formation assay and in vivo. This reversal in phenotype appears to enhance vessel formation and is a new form of signaling for Notch ligands that expands their signaling potential beyond cell-cell contact

Search for B0→π0π0B^{0}\to \pi^{0}\pi^{0} Decay

We have searched for the charmless hadronic decay of B0 mesons into two neutral pions. Using 9.13fb^-1 taken at the Upsilon(4S) with the CLEO detector, we obtain an improved upper limit for the branching fraction BR(B0-->pi0pi0) < 5.7*10^-6 at the 90% confidence level.Comment: pages postscript, also available through http://w4.lns.cornell.edu/public/CLN

Search for WHWH associated production in ppˉp \bar{p} collisions at s=1.96 TeV\sqrt{s}=1.96\,{\rm TeV}

This report describes a search for associated production of $W$ and Higgs bosons based on data corresponding to an integrated luminosity of $\cal{L}$$\approx 5.3 \rm fb^{-1}$ collected with the D0 detector at the Fermilab Tevatron $p\bar{p}$ Collider. Events containing a $W\rightarrow \ell \nu$ candidate (with $\ell$ corresponding to $e$ or $\mu$) are selected in association with two or three reconstructed jets. One or two of the jets are required to be consistent with having evolved from a $b$ quark. A multivariate discriminant technique is used to improve the separation of signal and backgrounds. Expected and observed upper limits are obtained for the product of the $WH$ production cross section and branching ratios and reported in terms of ratios relative to the prediction of the standard model as a function of the mass of the Higgs boson ($M_{H}$). The observed and expected 95% C.L. upper limits obtained for an assumed $M_{H}=115 \rm GeV$ are, respectively, factors of 4.5 and 4.8 larger than the value predicted by the standard model.Comment: 25 pages, 14 figure

Search for first generation leptoquark pair production in the electron + missing energy + jets final state

We present a search for the pair production of first generation scalar leptoquarks (LQ) in data corresponding to an integrated luminosity of 5.4 fb$^{-1}$ collected with the D0 detector at the Fermilab Tevatron Collider in ppbar collisions at $\sqrt{s}=1.96$ TeV. In the channel $LQ \bar{LQ} \rightarrow e\nu_e qq'$, where q, q' are u or d quarks, no significant excess of data over background is observed, and we set a 95% C.L. lower limit of 326 GeV on the leptoquark mass, assuming equal probabilities of leptoquark decays to eq and $\nu_e q'$.Comment: 7 pages, 6 figures, submitted to PRD-R

Zgamma production and limits on anomalous ZZgamma and Zgammagamma couplings in ppbar collisions at sqrt(s)=1.96 TeV

We present a measurement of ppbar->Zgamma->ll+gamma (l = e, mu) production with a data sample corresponding to an integrated luminosity of 6.2 fb^{-1} collected by the D0 detector at the Fermilab Tevatron ppbar Collider. The results of the electron and muon channels are combined, and we measure the total production cross section and the differential cross section dsigma/dp_T^gamma, where p_T^gamma is the momentum of the photon in the plane transverse to the beamline. The results obtained are consistent with the standard model predictions from next-to-leading order calculations. We use the transverse momentum spectrum of the photon to place limits on anomalous ZZgamma and Zgammagamma couplings

Search for a Narrow ttbar Resonance in ppbar Collisions at sqrt{s}=1.96 TeV

We report a search for a narrow ttbar resonance that decays into a lepton+jets final state based on an integrated luminosity of 5.3/fb of proton-antiproton collisions at sqrt{s}=1.96 TeV collected by the D0 Collaboration at the Fermilab Tevatron Collider. We set upper limits on the production cross section of such a resonance multiplied by its branching fraction to ttbar which we compare to predictions for a leptophobic topcolor Z' boson. We exclude such a resonance at the 95% confidence level for masses below 835 GeV.Comment: 7 pages, 3 figures, submitted to Physical Review Letter

Search for Zgamma events with large missing transverse energy in ppbar collisions at sqrt(s)=1.96 TeV

We present the first search for supersymmetry (SUSY) in Zgamma final states with large missing transverse energy using data corresponding to an integrated luminosity of 6.2 fb-1 collected with the D0 experiment in ppbar collisions at sqrt(s)=1.96 TeV. This signature is predicted in gauge-mediated SUSY-breaking models, where the lightest neutralino is the next-to-lightest supersymmetric particle (NLSP) and is produced in pairs, possibly through decay from heavier supersymmetric particles. The NLSP can decay either to a Z boson or a photon and an associated gravitino that escapes detection. We exclude this model at the 95% C.L. for SUSY breaking scales of Lambda < 87 TeV, corresponding to neutralino masses of < 151 GeV.Comment: submitted to Phys. Rev. Let