RESUMO
The Snail gene product is a transcriptional repressor of E-cadherin expression and an inducer of the epithelial-to-mesenchymal transition in several epithelial tumor cell lines. This report presents data indicating that Snail function is controlled by its intracellular location. The cytosolic distribution of Snail depended on export from the nucleus by a CRM1-dependent mechanism, and a nuclear export sequence (NES) was located in the regulatory domain of this protein. Export of Snail was controlled by phosphorylation of a Ser-rich sequence adjacent to this NES. Modification of this sequence released the restriction created by the zinc finger domain and allowed nuclear export of the protein. The phosphorylation and subcellular distribution of Snail are controlled by cell attachment to the extracellular matrix. Suspended cells presented higher levels of phosphorylated Snail and an augmented extranuclear localization with respect to cells attached to the plate. These findings show the existence in tumor cells of an effective and fine-tuning nontranscriptional mechanism of regulation of Snail activity dependent on the extracellular environment.
Assuntos
Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Receptores Citoplasmáticos e Nucleares , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transporte Ativo do Núcleo Celular/fisiologia , Animais , Sequência de Bases , Núcleo Celular/genética , Núcleo Celular/metabolismo , Citosol/metabolismo , Matriz Extracelular/metabolismo , Humanos , Carioferinas/metabolismo , Leucina/metabolismo , Camundongos , Dados de Sequência Molecular , Sinais de Localização Nuclear/genética , Sinais de Localização Nuclear/metabolismo , Fosforilação , Estrutura Terciária de Proteína , Transporte Proteico/fisiologia , Serina/metabolismo , Fatores de Transcrição da Família Snail , Frações Subcelulares , Células Tumorais Cultivadas , Proteína Exportina 1RESUMO
Photoperiod controls several responses throughout the plant life cycle, like germination, flowering, tuber formation, onset of bud dormancy, leaf abscission, and cambium activity. From these processes, flowering has been most extensively studied, especially in Arabidopsis thaliana. Photoperiod sensing by the function of photoreceptors and the circadian clock appears to regulate flowering time via Arabidopsis CONSTANS (AtCO), a putative transcription factor that accelerates flowering in response to long days. The genetic factors controlling plant photoperiodic responses other than flowering are little known. However, interspecific grafting experiments demonstrated that the flower-inducing (florigen) and tuber- inducing (tuberigen) signals are functionally exchangeable. Here we show that constitutive overexpression in potato of the Arabidopsis flowering-time gene AtCO impairs tuberization under short-day inductive conditions; AtCO overexpressing lines require prolonged exposure to short days to form tubers. Grafting experiments using these lines indicated that AtCO exerts its inhibitory effect on tuber formation by acting in the leaves. We propose that a conserved photoperiodic functional module may be involved in controlling distinct photoperiod-regulated evocation responses in different species. This module would involve the action of CONSTANS in the production of the elusive and long-distance acting florigen-tuberigen signal(s) in the leaves.
