RESUMO
The endoplasmic reticulum is traditionally perceived as the key compartment for regulating intracellular cholesterol metabolism. Increasing evidence suggests that the endocytic pathway provides an additional regulatory level governing intracellular cholesterol trafficking and homoeostasis. Sterols can enter, and apparently also exit, endosomal compartments via both vesicular and non-vesicular mechanisms. A number of studies have focused on endosomal sterol removal as its defects lead to cholesterol storage diseases. So far, the bulk of evidence on endosomal sterol egress describes the involvement of membrane trafficking machineries. Interestingly, two late endosomal sterol-binding proteins were recently shown to regulate the movement of late endosomes along cytoskeletal tracks. These studies provide the first indications of how non-vesicular and vesicular mechanisms may co-operate in endosomal sterol trafficking.
Assuntos
Colesterol/metabolismo , Endossomos/metabolismo , Vesículas Transportadoras/fisiologia , Animais , Membrana Celular/metabolismo , Humanos , Vesículas Transportadoras/química , Vesículas Transportadoras/metabolismoRESUMO
The prevention of flower formation is important for avoiding the spread of transgenes from genetically modified plants into wild populations. Moreover, the resources not expended for the generation of flowers and fruits might be allocated to increased vegetative growth. We have been developing methods for preventing flower formation in silver birch (Betula pendula), a tree species of considerable economical importance in the boreal region. Here we study the suitability of the promoter of BpFRUITFULL-LIKE1 (BpFULL1, formerly BpMADS5) for tissue-specific ablation of inflorescences in Arabidopsis, tobacco and birch. With all these species, the development of inflorescences was successfully prevented. The results show that the BpFULL1::BARNASE construct has potential biotechnological applications in different plant species.
Assuntos
Betula/genética , Flores/crescimento & desenvolvimento , Flores/genética , Inibidores do Crescimento/genética , Proteínas de Plantas/genética , Proteínas Recombinantes de Fusão/genética , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Bactérias , Betula/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia , Regiões Promotoras Genéticas/genética , Ribonucleases/genética , Nicotiana/genética , Nicotiana/crescimento & desenvolvimentoRESUMO
Cholesterol entering cells in low-density lipoproteins (LDL) via receptor-mediated endocytosis is transported to organelles of the late endocytic pathway for degradation of the lipoprotein particles. The fate of the free cholesterol released remains poorly understood, however. Recent observations suggest that late-endosomal cholesterol sequestration is regulated by the dynamics of lysobisphosphatidic acid (LBPA)-rich membranes [1]. Genetic studies have pinpointed a protein, Niemann-Pick C-1 (NPC-1), that is required for the mobilization of late-endosomal/lysosomal cholesterol by an unknown mechanism [2]. Here, we report the removal of accumulated cholesterol by overexpression of the NPC-1 protein in NPC-1-deficient fibroblasts from patients with Niemann-Pick disease, and in normal fibroblasts upon release of a progesterone-induced block of cholesterol transport. We show that late-endosomal/lysosomal cholesterol mobilization is specifically inhibited by microinjection of Rab GDP-dissociation inhibitor (Rab-GDI). Moreover, clearance of the cholesterol deposits by NPC-1 in patients' fibroblasts is accompanied by the redistribution of LBPA and of a lysosomal hydrolase that utilizes the mannose-6-phosphate receptor. Our results reveal, for the first time, the involvement of a specific molecular component of the membrane-trafficking machinery in cholesterol transport and the coupling of late-endosomal cholesterol egress to the trafficking of other lipid and protein cargo.
