RESUMEN
Cell-specific expression patterns of the Eucalyptus gunnii cinnamoyl coenzymeA reductase (EgCCR) and cinnamyl alcohol dehydrogenase (EgCAD2) promoters were analyzed by promoter-GUS histochemistry in the primary and secondary xylem tissues from floral stems and roots of Arabidopsis thaliana. Expression patterns indicated that the EgCCR and EgCAD2 genes were expressed in a coordinated manner in primary and secondary xylem tissues of the Arabidopsis floral stem and root. Both genes were expressed in all lignifying cells (vessel elements, xylem fibers and paratracheal parenchyma cells) of xylem tissues. The capacity for long-term monolignol production appeared to be related to the cell-specific developmental processes and biological roles of different cell types. Our results suggested that lignification of short-lived vessel elements was achieved by a two-step process involving (i) monolignol production by vessel elements prior to vessel programmed cell death and (ii) subsequent monolignol production by vessel-associated living paratracheal parenchyma cells following vessel element cell death. EgCCR and EgCAD2 gene expression patterns suggested that the process of xylem cell lignification was similar in both primary and secondary xylem tissues in Arabidopsis floral stems and roots.
Asunto(s)
Oxidorreductasas de Alcohol/biosíntesis , Aldehído Oxidorreductasas/biosíntesis , Arabidopsis , Eucalyptus/enzimología , Proteínas de Plantas/biosíntesis , Xilema/enzimología , Oxidorreductasas de Alcohol/genética , Aldehído Oxidorreductasas/genética , Arabidopsis/citología , Arabidopsis/enzimología , Arabidopsis/genética , Eucalyptus/citología , Eucalyptus/genética , Regulación Enzimológica de la Expresión Génica/genética , Regulación de la Expresión Génica de las Plantas/genética , Proteínas de Plantas/genética , Raíces de Plantas/citología , Raíces de Plantas/enzimología , Raíces de Plantas/genética , Tallos de la Planta/citología , Tallos de la Planta/enzimología , Tallos de la Planta/genética , Plantas Modificadas Genéticamente , Xilema/citología , Xilema/genéticaRESUMEN
In this report, we have shown that the yeast amphiphysin-like protein Rvs167p was localized mainly in small cortical patches throughout the cell in unbudding cells. During budding, the patches were polarized at bud emergence site. During mating, Rvs167p was concentrated at the tip of the shmoo. Rvs167p colocalized with actin patches during yeast vegetative growth and mating. Complete disruption of the actin cytoskeleton using Latrunculin-A did not affect Rvs167p localization in patches throughout the cell. In rvs167 mutant cells, actin patches are mislocalized and in rvs161 or abp1 mutant cells, Rvs167p localization is not affected. These observations suggest that Rvs167p may localize the actin cortical complex properly. Finally, the amphiphysin-conserved N-terminal domain of Rvs167p, called the BAR domain, was required but not sufficient for the correct localization of the protein.
Asunto(s)
Actinas/fisiología , Proteínas Fúngicas/fisiología , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/fisiología , Saccharomyces cerevisiae/ultraestructura , Actinas/análisis , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , División Celular , Citoesqueleto/efectos de los fármacos , Citoesqueleto/ultraestructura , Proteínas Fúngicas/análisis , Proteínas Fúngicas/genética , Genotipo , Proteínas Fluorescentes Verdes , Proteínas Luminiscentes/análisis , Toxinas Marinas/farmacología , Proteínas de Microfilamentos , Proteínas del Tejido Nervioso/fisiología , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes de Fusión/análisis , Saccharomyces cerevisiae/genética , Tiazoles/farmacología , TiazolidinasRESUMEN
Mutations in RVS161 and RVS167 yeast genes induce identical phenotypes associated to actin cytoskeleton disorders. The whole Rvs161 protein is similar to the amino-terminal part of Rvs167p, thus defining a RVS domain. In addition to this domain, Rvs167p contains a central glycine-proline-alanine rich domain and a SH3 domain. To assess the function of these different domains we have expressed recombinant Rvs proteins in rvs mutant strains. Phenotype analysis has shown that the RVS and SH3 domains are necessary for phenotypical complementation, whereas the GPA domain is not. Moreover, we have demonstrated that the RVS domains from Rvs161p and Rvs167p have distinct roles, and that the SH3 domain needs the specific RVS domain of Rvs167p to function. These results suggest that Rvs161p and Rvs167p play distinct roles, while acting together in a common function.
Asunto(s)
Proteínas del Citoesqueleto , Proteínas Fúngicas/fisiología , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/fisiología , Actinas/metabolismo , Sitios de Unión , Citoesqueleto/metabolismo , Proteínas Fúngicas/genética , Proteínas de Microfilamentos , Fenotipo , Proteínas Recombinantes de Fusión/metabolismo , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/crecimiento & desarrolloRESUMEN
The rvs161 and rvs167 mutant cells exhibit several identical phenotypes including sensitivity to several different growth conditions and morphological defects such as alteration of the actin cytoskeleton and budding patterns. The selection of genes that, when overexpressed, are able to suppress the reduced viability upon carbon starvation of the rvs167 mutant strain, has allowed the cloning of the SUR7 gene (Accession Number Z46729x11). We showed that the suppressive ability of the overexpressed SUR7 gene concerns all the rvs167 phenotypes. However, this suppression is only partial since the rvs167-suppressed strain is not of wild-type phenotype. Moreover, SUR7 is also able to suppress partially the phenotypes exhibited by the rvs161 and rvs167 and rvs161 mutant strains. The SUR7 gene encodes a putative integral membrane protein with four transmembrane domains. Furthermore, sequence comparisons revealed that Sur7p and two other proteins, Yn1194p and Yd1222p, present significant sequence and structural similarities. Taken together, these results strongly suggest that the Rvs161 and Rvs167 proteins act together in relation with Sur7p. Moreover, the putative transmembranous character of Sur7p suggests a membrane localization of the Rvs function, a localization which is consistent with the different rvs phenotypes and the actin-Rvs167p interaction.
Asunto(s)
Proteínas del Citoesqueleto , Proteínas Fúngicas/genética , Genes Fúngicos , Genes Supresores , Proteínas de la Membrana/genética , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Actinas/metabolismo , Secuencia de Aminoácidos , Clonación Molecular , Proteínas Fúngicas/fisiología , Proteínas de la Membrana/fisiología , Datos de Secuencia Molecular , MutaciónRESUMEN
The actin cytoskeleton cells is altered in rvs161 mutant yeast, with the defect becoming more pronounced under unfavorable growth conditions, as described for the rvs167 mutant. The cytoskeletal alteration has no apparent effect on invertase secretion and polarized growth. Mutations in RVS161, just as in RVS167, lead to a random budding pattern in a/alpha diploid cells. This behavior is not observed in a/a diploid cells homozygous for the rvs161-1 or rvs167-1 mutations. In addition, sequence comparisons revealed that amphiphysin, a protein first found in synaptic vesicles of chicken and shown to be the autoantigen of Stiff Man syndrome, presents similarity with both Rvs proteins. Furthermore, limited similarities with myosin heavy chain and tropomyosin alpha chain from higher eukaryotic cells allow for the definition of a possible consensus sequence. The finding of related sequences suggests the existence of a function for these proteins that is conserved among eukaryotic organisms.