RESUMEN
NEDD8 is an ubiquitin-like molecule that covalently binds to target proteins through an enzymatic cascade analogous to ubiquitylation. This modifier is known to bind to p53 and p73, as well as all Cullin family proteins, which are essential components of Skp1/Cul-1/F-box protein (SCF)-like Ub ligase complexes. Here, we focused on a genomic analysis of the genes involved in the NEDD8 conjugation pathway in Schistosoma mansoni. The results revealed seven genes related to NEDD8 conjugation that are conserved in Schistosoma japonicum, Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens. We performed quantitative RT-PCR (qRT-PCR), which showed differential profiles for Smnedd8, Smapp1, Smuba3, Smube2f, Smdcn1, Smrbx and Smsenp8 throughout the life cycle of S. mansoni. Upregulation was observed in 3-day-old schistosomula and adult worms for all analysed genes. We also analysed the transcription levels of Cullin family members Smp63 and Smp73, and observed upregulation in early schistosomula, while cercariae and adult worms showed expression levels similar to one another. Taken together, these results suggest that the NEDDylation/DeNEDDylation pathway controls important cellular regulators during worm development from cercariae to schistosomula and, finally, to adult.
Asunto(s)
Proteínas Cullin/metabolismo , Procesamiento Proteico-Postraduccional , Schistosoma mansoni/metabolismo , Enzimas Ubiquitina-Conjugadoras/genética , Ubiquitinas/genética , Secuencia de Aminoácidos , Animales , Biomphalaria , Proteínas Cullin/genética , Regulación de la Expresión Génica , Genómica , Proteínas del Helminto/genética , Proteínas del Helminto/metabolismo , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Filogenia , Estructura Terciaria de Proteína , ARN de Helminto/genética , Schistosoma mansoni/enzimología , Schistosoma mansoni/genética , Schistosoma mansoni/crecimiento & desarrollo , Esquistosomiasis mansoni/parasitología , Alineación de Secuencia , Transcriptoma , Enzimas Ubiquitina-Conjugadoras/metabolismo , Complejos de Ubiquitina-Proteína Ligasa/genética , Complejos de Ubiquitina-Proteína Ligasa/metabolismo , Ubiquitinas/metabolismoRESUMEN
Body measurement traits, influenced by genes and environmental factors, play numerous important roles in the value assessment of productivity and economy. There has been some indication that ANAPC13 influences adult height. We used PCR-SSCP and DNA sequencing technology to identify polymorphisms in the ANAPC13 gene. A polymorphism in intron 1 (A > G at base 17) was identified and an additional polymorphic site (C > T at base 42) was also uncovered, which accompanied the previous polymorphism in more than 98% of the subjects. The two novel polymorphisms in exon 1 were assayed and potential associations with body measurement traits were evaluated in 404 individuals. Three genotypes were detected in the study group, named AACC, AGCT and GGTT. Significant differences were observed between genotypes AACC and AGCT for body length, withers height, hip height, hip width, heart girth, pin bone width. However, no associations were found among any genotypes and chest depth. We conclude that polymorphisms and mutations in non-coding regions of the ANAPC13 gene significantly affect body measurement traits.
Asunto(s)
Pesos y Medidas Corporales , Bovinos/anatomía & histología , Bovinos/genética , Estudios de Asociación Genética , Polimorfismo de Nucleótido Simple/genética , Carácter Cuantitativo Heredable , Complejos de Ubiquitina-Proteína Ligasa/genética , Ciclosoma-Complejo Promotor de la Anafase , Animales , Secuencia de Bases , Distribución de Chi-Cuadrado , China , Exones/genética , Frecuencia de los Genes/genética , Sitios Genéticos/genética , Genotipo , Análisis de los Mínimos Cuadrados , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-Simple/genética , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: The orderly progression through mitosis is regulated by the Anaphase-Promoting Complex (APC), a large multiprotein E3 ubiquitin ligase that targets key cell-cycle regulators for destruction by the 26 S proteasome. The APC is composed of at least 11 subunits and associates with additional regulatory activators during mitosis and interphase cycles. Despite extensive research on APC and activator functions in the cell cycle, only a few components have been functionally characterized in plants. RESULTS: Here, we describe an in-depth search for APC subunits and activator genes in the Arabidopsis, rice and poplar genomes. Also, searches in other genomes that are not completely sequenced were performed. Phylogenetic analyses indicate that some APC subunits and activator genes have experienced gene duplication events in plants, in contrast to animals. Expression patterns of paralog subunits and activators in rice could indicate that this duplication, rather than complete redundancy, could reflect initial specialization steps. The absence of subunit APC7 from the genome of some green algae species and as well as from early metazoan lineages, could mean that APC7 is not required for APC function in unicellular organisms and it may be a result of duplication of another tetratricopeptide (TPR) subunit. Analyses of TPR evolution suggest that duplications of subunits started from the central domains. CONCLUSIONS: The increased complexity of the APC gene structure, tied to the diversification of expression paths, suggests that land plants developed sophisticated mechanisms of APC regulation to cope with the sedentary life style and its associated environmental exposures.
Asunto(s)
Evolución Molecular , Proteínas de Plantas/genética , Plantas/genética , Complejos de Ubiquitina-Proteína Ligasa/genética , Secuencia de Aminoácidos , Ciclosoma-Complejo Promotor de la Anafase , Arabidopsis/genética , Secuencia de Bases , Chlorophyta/enzimología , Chlorophyta/genética , Mapeo Cromosómico , Cromosomas de las Plantas/genética , Regulación de la Expresión Génica de las Plantas , Variación Genética , Genoma de Planta/genética , Datos de Secuencia Molecular , Oryza/genética , Filogenia , Proteínas de Plantas/clasificación , Plantas/enzimología , Populus/genética , Subunidades de Proteína/clasificación , Subunidades de Proteína/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rhodophyta/enzimología , Rhodophyta/genética , Especificidad de la Especie , Sintenía , Complejos de Ubiquitina-Proteína Ligasa/clasificaciónRESUMEN
Sister-chromatid separation and exit from mitosis require ubiquitin-mediated proteolysis of cell cycle regulators such as cyclin B and securin. The specificity of the reaction is controlled by an ubiquitin-ligase multiprotein complex known as APC (Anaphase Promoting Complex). Comparison of the coding sequences of Arabidopsis genes with the Genbank database reveals extensive homology of the predicted ORFs with the corresponding proteins of other eukaryotes, indicating that the APC is well conserved in plants. However, different from other eukaryotes, the Arabidopsis genes have some particular characteristics, such as the presence of two copies of the CDC27 gene. Furthermore, expression analyses of the AtAPC genes disclose complex profiles that differ, depending on the tissue examined. In actively dividing cell suspensions there is a direct correspondence between the rates of proliferation and mRNA levels from the AtAPC components. On the other hand, in plant organs, dark-grown seedlings and during leaf growth, this correlation is lost and the AtAPC genes are highly expressed in tissues with low overall cell division. Moreover, expression patterns diverge between the subunit genes, raising the possibility that there could be more than one form of the APC, which would execute distinct functions during plant development. The results suggest that an important layer of regulation of APC/C in plants could operate through subunit availability in specific tissues and/or cellular compartments.