Autor(es): González-Calixto Cecilia, Cázares-Raga Febe E, Cortés-Martínez Leticia, Del Angel Rosa María, Medina-Ramírez Fern-o, Mosso Clemente, Ocádiz-Ruiz Ramón, Valenzuela Jesús G, Rodríguez Mario Henry, Hernández-Hernández Fidel de la Cruz

Resumo: The Receptor for Activated C Kinase 1 (RACK1), a scaffold protein member of the tryptophan-aspartate (WD) repeat family, folds in a seven-bladed ?-propeller structure that permits the association of proteins to form active complexes. Mosquitoes of the genus Aedes sp., are vectors of virus producing important diseases such as: dengue, chikungunya - yellow fever. Based on the highly conserved gene sequence of AeaeRACK1 of the mosquito Aedes aegypti we characterized the mRNA - protein of the homologous AealRACK1 from the Ae. albopictus-derived cell line C6/36 HT. Two protein species differing in MW/pI values were observed at 35kDa/8.0 - 36kDa/6.5. The behavior of AealRACK1 was studied inducing stress with serum deprivation - the glucocorticoid dexamethasone. Both stressors induced increase of the expression of AealRACK1 mRNA - proteins. In serum-deprived cells AealRACK1 protein was located cortically near the plasma membrane in contrast to dexamethasone-treated cells where the protein formed a dotted pattern in the cytoplasm. In addition, 33 protein partners were identified by immunoprecipitation - mass spectrometry. Most of the identified proteins were ribosomal, involved in signaling pathways - stress responses. Our results suggest that AealRACK1 in C6/36 HT cells respond to stress increasing its synthesis - producing phosphorylated activated form. Insect cells adapt to numerous environmental stressors, including chemicals - invasion of pathogenic microorganisms among others, coordinating cellular - organismal responses. Individual cells sense the environment using receptors that trigger signaling pathways that regulate expression of specific effector proteins -/or cellular responses as movement or secretion. In the coordination of responses to stress, scaffold proteins are pivotal molecules that recruit other proteins forming active complexes. The Receptor for Activated C Kinase 1 (RACK1) is the best studied member of the conserved tryptophan-aspartate (WD) repeat family. RACK1 folds in a seven-bladed ?-propeller structure - it could be activated during stress, participating in different signaling pathways. The presence - activities of RACK1 in mosquitoes had not been documented before, in this work the molecule is demonstrated in an Aedes albopictus-derived cell line - its reaction to stress is observed under the effect of serum deprivation - the presence of glucocorticoid analog dexamethasone, a chemical used to cause stress in vitro.

Palavras-Chave: Aedes mosquito; C6/36 HT cells; Receptor for Activated C Kinase (RACK1); Scaffold protein; Stress conditions

Imprenta: Journal of Proteomics, v. 119, p. 45-60, 2015

Identificador do objeto digital: 10.1016/j.jprot.2014.11.019

Descritores: Chikungunya virus - Biosynthesis ; Chikungunya virus - Cell ; Chikungunya virus - Molecular structure ; Chikungunya virus - Protein synthesis ; Chikungunya virus - Proteins ; Chikungunya virus - Infectious diseases ; Chikungunya virus - Viral infections ; Chikungunya Virus - Virus ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue

Data de publicação: 2015

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