{"id":116201,"date":"2018-03-11T10:45:07","date_gmt":"2018-03-11T10:45:07","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/control-diferencial-de-la-ruta-de-map-quinasas-de-integridad-celular-por-gtpasas-de-la-familia-rho-en-schizosaccharomyces-pombe\/"},"modified":"2018-03-11T10:45:07","modified_gmt":"2018-03-11T10:45:07","slug":"control-diferencial-de-la-ruta-de-map-quinasas-de-integridad-celular-por-gtpasas-de-la-familia-rho-en-schizosaccharomyces-pombe","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/microbiologia\/control-diferencial-de-la-ruta-de-map-quinasas-de-integridad-celular-por-gtpasas-de-la-familia-rho-en-schizosaccharomyces-pombe\/","title":{"rendered":"Control diferencial de la ruta de map quinasas de integridad celular por gtpasas de la familia rho en schizosaccharomyces pombe"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Laura Sanchez Mir <\/strong><\/h2>\n<p>Resumen las rutas de se\u00f1alizaci\u00f3n mediadas por map quinasas (mapk) tienen un papel fundamental en la respuesta de c\u00e9lulas eucariotas frente a est\u00edmulos ambientales. La levadura con fisi\u00f3n schizosaccharomyces pombe representa un modelo particularmente adecuado para abordar estudios b\u00e1sicos sobre dichas rutas de se\u00f1alizaci\u00f3n, debido a la gran homolog\u00eda funcional que existe entre sus circuitos reguladores y los de organismos superiores. En s. Pombe, la ruta de mapks de integridad celular (cip) participa en la regulaci\u00f3n de numerosos procesos biol\u00f3gicos como la construcci\u00f3n de la pared celular, la citocinesis, la morfog\u00e9nesis, la fusi\u00f3n de vacuolas durante el estr\u00e9s hipot\u00f3nico, y la homeostasis i\u00f3nica mediante la modulaci\u00f3n de la activaci\u00f3n de la mapk pmk1 en respuesta a cambios en las condiciones ambientales.  aunque la gtpasa rho2 es el principal activador de la ruta de integridad celular a trav\u00e9s del ort\u00f3logo de pkc pck2, pmk1 puede ser activada en ausencia de ambos dependiendo de la naturaleza del est\u00edmulo recibido, lo que indica la existencia de elementos reguladores adicionales aguas arriba del m\u00f3dulo de mapks. Se ha propuesto a la gtpasa rho1 como candidata para tal funci\u00f3n, mientras que el ort\u00f3logo de pkc pck1 parece regular negativamente la actividad de la cip. Hemos investigado el posible papel de rho1 y pck1 como reguladores de la cip empleando un mutante hipoactivo de rho1 (rho1-596) y un mutante carente de pck1 en distintos fondos gen\u00e9ticos. Las evidencias obtenidas mostraron que rho1 y pck1 son activadores de la ruta adicionalmente a rho2 y pck2, lo que pone de manifiesto la naturaleza ramificada de los mecanismos de se\u00f1alizaci\u00f3n que act\u00faan aguas arriba del m\u00f3dulo de mapks. por otra parte rho2 se localiza en la membrana plasm\u00e1tica y en el \u00e1rea de divisi\u00f3n celular, y es farnesilada in vivo en el motivo -caax de su extremo c-terminal, siendo esta modificaci\u00f3n esencial para su localizaci\u00f3n y funci\u00f3n activadora de la cip. Sin embargo, la presencia de un residuo de ciste\u00edna previo al motivo -caax sugiere que rho2 podr\u00eda encontrarse palmitoilada en dicho residuo. Para estudiar la relevancia biol\u00f3gica de la palmitoilaci\u00f3n de rho2 creamos una serie de cepas que expresan versiones mutantes tanto de rho2 como de rho1 afectadas en lipidaci\u00f3n, y estudiamos su localizaci\u00f3n y funci\u00f3n biol\u00f3gica. Los resultados obtenidos permitieron concluir que en la levadura con fisi\u00f3n, la gtpasa rho2 es palmitoilada in vivo, siendo esta modificaci\u00f3n cr\u00edtica para su correcta localizaci\u00f3n en membrana plasm\u00e1tica y para ejercer su regulaci\u00f3n de la morfog\u00e9nesis y la ruta de integridad celular.  por \u00faltimo, son numerosos los estudios que han puesto de manifiesto la importancia de la localizaci\u00f3n subcelular de los componentes de los m\u00f3dulos de map quinasas en la respuesta a estr\u00e9s. En el caso de la ruta de integridad celular, la mapk pmk1 muestra una localizaci\u00f3n n\u00facleo\/citoplasm\u00e1tica constitutiva, as\u00ed como en el huso mit\u00f3tico, el corp\u00fasculo polar del huso, y en el septo durante la separaci\u00f3n celular. Sin embargo, a diferencia del modelo descrito en erk1\/2 en mam\u00edferos, el patr\u00f3n de localizaci\u00f3n de pmk1 no se modifica en respuesta a estr\u00e9s, lo que indica que tanto la forma activa como inactiva de la map quinasa son capaces de entrar en el n\u00facleo. En base a esto, nos planteamos el estudio de la relevancia biol\u00f3gica de la localizaci\u00f3n nuclear de pmk1 sobre su funci\u00f3n biol\u00f3gica. Para ello recurrimos a la caracterizaci\u00f3n de mutantes de s. Pombe que expresan una versi\u00f3n de pmk1 anclada a la membrana plasm\u00e1tica y excluida del n\u00facleo constitutivamente en distintos fondos gen\u00e9ticos. Los resultados mostraron que aunque la localizaci\u00f3n nuclear de pmk1 en schizosaccharomyces pombe no es cr\u00edtica para ejercer la mayor parte de sus funciones biol\u00f3gicas, s\u00ed es necesaria para la regulaci\u00f3n transcripcional en respuesta al estr\u00e9s de pared celular, lo que revela la importancia del control espacio-temporal de la actividad de las mapks en los organismos eucariotas.   summary mitogen-activated protein kinase (mapk) signaling pathways play a fundamental role in the response of eukaryotic cells to environmental changes. Fission yeast schizosaccharomyces pombe has emerged as an excellent model organism to study mechanisms and cellular events linked to mapk activation, given the significant functional homology between their regulatory circuits and those of higher cells. In s. Pombe the cell integrity pathway (cip) participates in multiple biological processes like cell wall construction, vacuole fusion during hypotonic stress, cytokinesis, morphogenesis, and ionic homeostasis by fine tuning of mapk pmk1 activation in response to various environmental conditions.  although the small gtpase rho2 is a main positive regulator operating upstream the cip through protein kinase c ortholog pck2, pmk1 can still be activated in the absence of either rho2 or pck2 depending on the nature of the stimulus, suggesting the existence of additional upstream regulatory elements to modulate its activity. The essential gtpase rho1 is a candidate to control the activity of the cip, whereas pck1 appears to negatively regulate pmk1 activity. To study the role of rho1 gtpase and pkc ortholog pck1 in the cip, we characterized a s. Pombe mutant that expresses hypomorphic rho1 allele (rho1-596) and a mutant lacking pck1 in distinct genetic backgrounds. The evidences obtained in this study revealed that rho1 and pck1 are positive upstream activators of the cip in addition to rho2 and pck2 during cell growth and cell wall stress. This model shows the branched nature of the upstream signal network that regulates the cip. rho2 positively regulates the cip and is involved in the control of cell polarity and cell wall biosynthesis in fission yeast. Rho2 locates along the plasma membrane and the division area and it has been described that becomes farnesylated in vivo at its -caax motif. This lipid modification appears essential for both localization and function within the mapk pathway. Moreover, there is a second conserved cysteine residue upstream the rho2 caax motif which might be palmitoylated in vivo like other rho gtpases such as human rhob. To investigate the role of palmitoylation in rho2-signaling we constructed rho2? Strains expressing the wild type, unpalmitoylated, or unprenylated versions of rho2 as well as rho2 chimeras fused to the carboxyl end from the essential gtpase rho1, and we study their location and rho2-dependent signalling within the cip. Our results showed that in fission yeast rho2 becomes palmitoylated in vivo and this modification is required for plasma membrane localization and proper signaling to the cip. pmk1 is constitutively localized in both cytoplasm and nucleus, as well as in the mitotic spindle and septum during cytokinesis. However, contrary to the current model for erk1\/2, its localization pattern appears unaffected by its activation status. In this context, we questioned the biological significance of nuclear localization of mapk pmk1. We have addressed this issue by characterization of mutants expressing pmk1 versions excluded from the cell nucleus and anchored to the plasma membrane in different genetic backgrounds. We have found that nuclear localization of pmk1 is not critical to exert their biological functions, although its presence in the nucleus is necessary for transcriptional regulation under cell wall stress. These evidences highlight the relevance of the spatial\/temporal activity control of mapk activity in eukaryotes.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Control diferencial de la ruta de map quinasas de integridad celular por gtpasas de la familia rho en schizosaccharomyces pombe<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Control diferencial de la ruta de map quinasas de integridad celular por gtpasas de la familia rho en schizosaccharomyces pombe <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Laura Sanchez Mir <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Murcia<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 24\/07\/2014<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Teresa Soto Pino<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: Mar\u00eda  senena Corbalan garcia <\/li>\n<li>Rafael Rodr\u00edguez daga (vocal)<\/li>\n<li>humberto Mart\u00edn brieva (vocal)<\/li>\n<li>Miguel angel Rodriguez (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Laura Sanchez Mir Resumen las rutas de se\u00f1alizaci\u00f3n mediadas por map quinasas (mapk) tienen un papel fundamental [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center 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