Mechanisms of spindle positioning during "Caenorhabditis elgans" asymmetric cell division

Résumé : Le positionnement correct du fuseau mitotique est crucial pour les divisions cellulaires asymétriques, car il gouverne le contrôle spatial de la division cellulaire et assure la ségrégation adéquate des déterminants cellulaires. Malgré leur importance, les mécanismes contrôlant le positionnement du fuseau mitotique sont encore mal compris. Chez l'embryon au stade une-cellule du nématode Caenorhabditis elegans, le fuseau mitotique est positionné de manière asymétrique durant l'anaphase grâce à l'action de générateurs de force situés au cortex cellulaire, et dont la nature était jusqu'alors indéterminée. Ces générateurs de force corticaux exercent une traction sur les microtubules astraux et sont dépendants de deux protéines Gα et de leurs protéines associées. Cette thèse traite de la nature de la machinerie responsable pour la génération des forces de tractions, ainsi que de son lien avec les protéines Gα et associées. Nous avons combiné des expériences de coupure par faisceau laser du fuseau mitotique avec le contrôle temporel de l'inactivation de gènes ou de l'exposition à des produits pharmacologiques. De cette manière, nous avons établi que la dynéine, un moteur se déplaçant vers l'extrémité négative des microtubules, ainsi que la dynamique des microtubules, sont toutes deux requises pour la génération efficace des forces de tractions. Nous avons démontré que les protéines Gα et leurs protéines associées GPR-1/2 et LIN-5 interagissent in vivo avec LIS-1, un composant du complexe de la dynéine. De plus, nous avons découvert que les protéines Gα, GPR-1/2 et LIN-5 promeuvent la présence du complexe de la dynéine au cortex cellulaire. Nos résultats suggèrent un mécanisme par lequel les protéines Gα permettent le recrutement cortical de GPR-1/2 et LIN-5, assurant ainsi la présence de la dynéine au cortex. Conjointement avec la dynamique des microtubules, ce mécanisme permet la génération des forces de tractions afin d'obtenir une division cellulaire correcte. Comme les mécanismes contrôlant le positionnement du fuseau mitotique et les divisions cellulaires asymétriques sont conservés au cours de l'évolution, nous espérons que les mécanismes élucidés par ce travail sont d'importance générale pour la génération de la diversité cellulaire durant le développement. De plus, ces mécanismes pourraient être applicables à d'autres divisions asymétriques, comme celle des cellules souches, dont le disfonctionnement peut entraîner la génération de cellules cancéreuses. Abstract : Proper spindle positioning is crucial for asymmetric cell division, because it controls spatial aspects of cell division and the correct inheritance of cell-fate determinants. However, the mechanisms governing spindle positioning remain incompletely understood. In the Caenorhabditis elegans one-cell stage embryo, the spindle becomes asymmetrically positioned during anaphase through the action of as-yet unidentified cortical force generators that pull on astral microtubules and that depend on two Gα proteins and associated proteins. This thesis addresses the nature of the force generation machinery and the link with the Gα and associated proteins. By performing spindle-severing experiments following temporally restricted gene inactivation and drug exposure, we established that microtubule dynamics and the minus-end directed motor dynein are both required for generating efficient pulling forces. We discovered that the Gα proteins and their associated proteins GPR-1/2 and LIN-5 interact in vivo with LIS-1, a component of the dynein complex. Moreover, we uncovered that LIN-5, GPR-1/2 and the Gα proteins promote the presence of the dynein complex at the cell cortex. Our findings suggest a mechanism by which the Gα proteins enable GPR-1/2 and LIN-5 recruitment to the cortex, thus ensuring the presence of cortical dynein. Together with microtubule dynamics, this allows pulling forces to be exerted and proper cell division to be achieved. Because the mechanisms of spindle positioning and asymmetric cell division are conserved across evolution, we expect the underlying mechanism uncovered here to be of broad significance for the generation of cell diversity during development. Moreover, this mechanism could be relevant for other asymmetric cell divisions, such as stem cell divisions, whose dysfunction may lead to the generation of cancer cells.

Report on the IowaCare program administered by the Iowa Medicaid Enterprise, a division of the Department of Human Services (DHS-IME), for the period July 1, 2005 through June 30, 2009

Report on the IowaCare program administered by the Iowa Medicaid Enterprise, a division of the Department of Human Services (DHS-IME), for the period July 1, 2005 through June 30, 2009

Wireless E911 Emergency Communications Fund Iowa Homeland Security and Emergency Management Division of the Iowa Department of Public Defense independent auditor's reports, basic financial statements and supplementary information, June 30, 2009.

Audit report on the Wireless E911 Emergency Communication Fund of the Iowa Homeland Security and Emergency Management Division of the Iowa Department of Public Defense for the year ended June 30, 2009

Iowa Insurance Division Report to the Governor for Year End 2001

Iowa Insurance Division Report to the Governor for Year End 2001

Iowa Insurance Division Report to the Governor for Year End 2002

Iowa Insurance Division Report to the Governor for Year End 2003

Iowa Insurance Division Report to the Governor for Year End 2003

Iowa Insurance Division Report to the Governor for Year End 2003

Iowa Insurance Division Report to the Governor for Year End 2004

Iowa Insurance Division Report to the Governor for Year End 2004

Iowa Insurance Division Report to the Governor for Year End 2005

Iowa Insurance Division Report to the Governor for Year End 2005

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Iowa Insurance Division Report to the Governor for Year End 2006

Iowa Insurance Division Report to the Governor for Year End 2007

Iowa Insurance Division Report to the Governor for Year End 2007

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Iowa Insurance Division Report to the Governor for Year End 2008

Report of recommendations to the Iowa Department of Education, Division of Vocational Rehabilitation Services, June 30, 2009

Report on the Iowa Department of Education, Division of Vocational Rehabilitation Services for the year ended June 30, 2009

Iowa Alcoholic Beverages Division, Annual Report, 2010

Iowa Alcoholic Beverages Division, Annual Report 2010

Division of Soil Conservation Soil and Water Quality Review, 2009

Special Points of Interest: • The Division of Soil Conservation celebrated its 70th anniversary July 1, 2009. The Iowa Soil Conservation: Laws were enacted in 1939 creating the state soil conservation agency and governing committee and providing for the creation of Iowa’s 100 soil and water conservation districts. • The Mines & Minerals Bureau, through the federal Abandoned Mine Land (AML) Program, worked with various watershed groups to again secure an additional $1 million dollars in funding for the construction on projects in Marion, Mahaska and Monroe Counties. • Iowa hosted the Mississippi River/Gulf of Mexico Hypoxia Task Force tour and meeting in September 2009.