Leukemia-associated RhoGEF (LARG) is a novel RhoGEF in cytokinesis and required for the proper completion of abscission.

Proper completion of mitosis requires the concerted effort of multiple RhoGEFs. Here we show that leukemia-associated RhoGEF (LARG), a RhoA-specific RGS-RhoGEF, is required for abscission, the final stage of cytokinesis, in which the intercellular membrane is cleaved between daughter cells. LARG colocalizes with α-tubulin at the spindle poles before localizing to the central spindle. During cytokinesis, LARG is condensed in the midbody, where it colocalizes with RhoA. HeLa cells depleted of LARG display apoptosis during cytokinesis with unresolved intercellular bridges, and rescue experiments show that expression of small interfering RNA-resistant LARG prevents this apoptosis. Moreover, live cell imaging of LARG-depleted cells reveals greatly delayed fission kinetics in abscission in which a population of cells with persistent bridges undergoes apoptosis; however, the delayed fission kinetics is rescued by Aurora-B inhibition. The formation of a Flemming body and thinning of microtubules in the intercellular bridge of cells depleted of LARG is consistent with a defect in late cytokinesis, just before the abscission event. In contrast to studies of other RhoGEFs, particularly Ect2 and GEF-H1, LARG depletion does not result in cytokinetic furrow regression nor does it affect internal mitotic timing. These results show that LARG is a novel and temporally distinct RhoGEF required for completion of abscission

Exile Vol. XLVI No. 1

44th Year Title Page 3 Epigraph by Ezra Pound 5 Statement of Policy 6 Table of Contents 7 Contributors Notes 37 Editorial Board 38 ART Incredible Lines by David Tulkin \u2701 8 Untitled by David Tulkin \u2701 12 Untitled by Patrick Yingling \u2703 18 Sculpted Body by David Tulkin \u2701 20 Untitled by Tanya Sheremeta \u2701 25 Untitled by David Tulkin \u2701 28 Untitled by Dena Behi \u2701 36 POETRY Wednesday by Mary Ann T. Davis \u2700 9 Music by Jessica Kramer \u2703 10-11 Worship During the Rainy Season by Allison Armbrister \u2701 19 Mercy by Mary Ann T. Davis \u2700 26-27 Albino Lizard by Matthew Martz \u2702 29 Filling of Lake Cumberland, 1951 by Allison Armbrister \u2701 35 PROSE Storm Drain by Matthew Martz \u2702 13-17 Creases by Stephanie M. Vaccaro \u2701 21-24 When It Rains by Matthew Martz \u2702 30-34 Statement of Policy This semester Exile instituted a new policy limiting submissions to those not involved in the production of the magazine. As always, all submissions are reviewed on an anonymous basis, and all editorial decisions are shared equally among the members of the Editorial Board. -6 Cover Art Untitled by Dena Behi \u2701 / Back Cover Art Untitled by Tanya Sheremeta \u2701 -38 Printed by Printing Arts Press -38 Matthew Martz, Albino Lizard, Storm Drain and When It Rains , redacted due to copyright restrictions

Exile Vol. XLVIII

46th Year Title Page 3 Epigraph by Ezra Pound 5 Table of Contents 7 Contributors\u27 Notes 59-60 Editorial Board 61 ART Connected by Christin Faison \u2704 8 Marcya by Harper Leich \u2704 12 Untitled by Derek Mong \u2704 19 Epistemological Torment by Matthew Sove \u2704 21 Consummatum Est I by Matthew Sove \u2704 27 Consummatum Est III by Matthew Sove \u2704 29 The Sheraton by Harper Leich \u2704 39 Untitled by Roman Sehling \u2703 43 My War by Matthew Sove \u2704 53 POETRY Chores, Then and Now by Dan Rohrer \u2704 9 Heaven by Chris Million \u2702 10-11 Loving her in Balinese by Ginna Fuselier \u2703 20 The Shower Room by Derek Mong \u2704 28 The Wailua by Jenny Silva \u2702 40-41 Good to the Last Drop by Chrissy Swinko \u2703 42 PROSE The House of Spider Webs by Dianna Craig \u2703 13-18 The Golem by Owen McGrann \u2703 22-26 All the String in Cincinnati by Rachel Colina \u2702 30-38 Passing by Matthew Martz \u2702 44-52 True War Story by Dianna Craig \u2703 54-58 All submissions are reviewed on an anonymous basis, and all editorial decisions are shared equally among the members of the Editorial Board. -61 Cover Art Out to Dry by Thomas DeCarlo \u2705 / Back Cover Art Untitled by Thomas DeCarlo \u2705 -61 Printing by Printing Arts Press -6

The experimental power of FR900359 to study Gq-regulated biological processes.

Despite the discovery of heterotrimeric αβγ G proteins ∼25 years ago, their selective perturbation by cell-permeable inhibitors remains a fundamental challenge. Here we report that the plant-derived depsipeptide FR900359 (FR) is ideally suited to this task. Using a multifaceted approach we systematically characterize FR as a selective inhibitor of Gq/11/14 over all other mammalian Gα isoforms and elaborate its molecular mechanism of action. We also use FR to investigate whether inhibition of Gq proteins is an effective post-receptor strategy to target oncogenic signalling, using melanoma as a model system. FR suppresses many of the hallmark features that are central to the malignancy of melanoma cells, thereby providing new opportunities for therapeutic intervention. Just as pertussis toxin is used extensively to probe and inhibit the signalling of Gi/o proteins, we anticipate that FR will at least be its equivalent for investigating the biological relevance of Gq

Ceremonies and Time in Shakespeare

This essay considers some moments in Shakespeare's texts which exemplify the Janus-faced quality of ceremonies: their enactment in the present looking backwards to past traditions and forwards to inaugurate new social relations. The argument draws on Victor Turner's theorization of ritual as an event that gives shape to “liminality,” that which “eludes or slips through the network of classification that normally locate states and positions in cultural space,” and argues that this applies to time as well. It also considers the construction of time in terms of kairos, a moment of time infused with meaning. The essay analyses ceremony in three Shakespearean genres. First, it examines Bertram's and Helena's ring exchange in All's Well That Ends Well as a “distended” ritual that collapses time. It then turns to Richard III, unpacking its complex sequence of ceremonies of betrothal, mourning, and sovereignty that are “continuously disrupted”. The final section describes the ceremonial time of romance in The Winter's Tale, unfolding the power invested in the kairotic time evoked by the oracle of Delphi, the sheep-shearing ceremony, and Paulina's “resurrection” of Hermione

Safety and Effectiveness of Meropenem in Infants With Suspected or Complicated Intra-abdominal Infections

Background. Intra-abdominal infections are common in young infants and lead to significant morbidity and mortality. Meropenem is a broad-spectrum antimicrobial with excellent activity against pathogens associated with intra-abdominal infections. The purpose of this study was to determine the safety and effectiveness of meropenem in young infants with suspected or complicated intra-abdominal infections

Master of Science

thesisMajor and trace element analyses and mineralogical data are presented for selected Plio-Pleistocene tuffs from the Shungura Formation which are useful in documenting hominid and vertebrate evolution in East Africa. The mainly fluvially deposited, hydrated tuffs are indicated as being comendites and pantellerites in all respects except for low Na20 and K20 values for which leaching is shown to be responsible. Mineral assemblages uncommon among conmedites and pantellerties are portrayed by the occurrence of chevkinite, manganiferous ilmenite, orthopyroxene, and plagioclase. Separating the tuffs into genetically related groups is difficult, due to similar trace element abundances and the evolutionary nature of volcanic chemistries. Similar chemistries may indicate that they all originate from the same general source area. Correlations on chemical bases are demonstrated between tuffs from the Shungura Formation and tuffs from both the Usno and Koobi Fora formations which are situated nearby in the Lake Turkana (formerly Lake Rudolf) area. The proposed source area of these tuffs is near Soddu on the western margin of the Ethiopian Rift. This was concluded from the following onsiderations: 1) the Plio-Pleistocene peralkaline rocks of the Ethiopian Rift are chemically and mineralogically very simila to the Shungura tuffs, and 2) tuffs common to both the Shungura and Koobi Fora Formations must have a source that is accessible to both the drainages responsible for transporting these tuffs to their present location

Leukemia-associated RhoGEF (LARG) is a novel RhoGEF in mitosis and required for the proper completion of abscission

Proper completion of mitosis requires the concerted effort of multiple RhoGEFs. Here we show that leukemia-associated RhoGEF (LARG), a RhoA specific RGS-RhoGEF, is required for abscission, the final stage of cytokinesis wherein the intercellular membrane is cleaved between daughter cells. LARG co-localizes with \u26agr;-tubulin at the spindle poles before localizing to the central spindle. During cytokinesis, LARG is condensed in the midbody where it colocalizes with RhoA. HeLa cells depleted of LARG display apoptosis during cytokinesis with unresolved intercellular bridges, and rescue experiments show that expression of siRNA-resistant LARG prevented this apoptosis. LARG is phosphorylated in early mitosis through a Cdk1-dependent pathway and is dephosphorylated upon anaphase transition. With the onset of cytokinesis, LARG translocates to membrane-associated structures and interacts with contractile ring components. Live cell imaging of LARG-depleted cells revealed greatly delayed fission kinetics in abscission wherein a population of cells with persistent bridges undergo apoptosis; however, these delayed fission kinetics appear to be rescued by Aurora B inhibition. The formation of a Flemming body and thinning of microtubules in the intercellular bridge of cells depleted of LARG is consistent with a defect in late cytokinesis, just prior to the abscission event. In contrast to studies of other RhoGEFs, particularly ECT2 and GEF-H1, LARG depletion does not result in cytokinetic furrow regression nor does it affect internal mitotic timing. Further, mutation of the conserved tyrosine residue in the DH core of LARG results in an inability to bind RhoA and apoptosis. These results establish LARG as a novel and temporally distinct RhoGEF required for the completion of abscission and mitotic viability. Moreover, the importance of LARG just prior to abscission suggests RhoA is critical downstream of contractile ring ingression in a secondary activation event.