RESUMO
Toll-like receptors (TLRs) trigger the innate immune system by responding to specific components of microorganisms. MyD88 and TRIF are Toll/interleukin (IL)-1 (TIR)-domain containing adapters, which play essential roles in TLR-mediated signalling via the MyD88-dependant and -independent pathways, respectively. Genes encoding several TLRs have been identified in the chicken genome, however, elements of their signalling pathways have not been well characterized. Here we describe the cloning of chicken MyD88 and TRIF orthologs, and examine the spatial and temporal expression of these genes. The chicken MyD88 cDNA was shown to have an open reading frame (ORF) of 1104 bp, encoding a predicted protein sequence of 368 aa, 8 aa short of a previously published coding sequence due to a premature stop codon. MyD88 gene expression was detected in each tissue tested except in muscle. The chicken TRIF cDNA possessed an ORF of 2205 bp, encoding a predicted protein sequence of 735 aa, which shared 37.3% similarity and 28.9% identity to human TRIF protein sequence. TRIF was ubiquitously expressed in all tissues.
Assuntos
Proteínas Adaptadoras de Transporte Vesicular/genética , Galinhas/genética , Fator 88 de Diferenciação Mieloide/genética , Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Sequência de Aminoácidos , Animais , Galinhas/metabolismo , Clonagem Molecular , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Dados de Sequência Molecular , Fator 88 de Diferenciação Mieloide/metabolismo , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Homologia de Sequência de Aminoácidos , Distribuição TecidualRESUMO
The small ubiquitin-related modifiers (SUMOs) are evolutionarily conserved polypeptides that are covalently conjugated to protein targets to modulate their subcellular localization, half-life, or activity. Steady-state SUMO conjugation levels increase in response to many different types of environmental stresses, but how the SUMO system is regulated in response to these insults is not well understood. Here, we characterize a novel mode of SUMO system control: in response to elevated alcohol levels, the Saccharomyces cerevisiae SUMO protease Ulp1 is disengaged from its usual location at the nuclear pore complex (NPC) and sequestered in the nucleolus. We further show that the Ulp1 region previously demonstrated to interact with the karyopherins Kap95 and Kap60 (amino acids 150 to 340) is necessary and sufficient for nucleolar targeting and that enforced sequestration of Ulp1 in the nucleolus significantly increases steady-state SUMO conjugate levels, even in the absence of alcohol. We have thus characterized a novel mechanism of SUMO system control in which the balance between SUMO-conjugating and -deconjugating activities at the NPC is altered in response to stress via relocalization of a SUMO-deconjugating enzyme.