RESUMO
The endoplasmic reticulum (ER) contains both cisternal and reticular elements in one contiguous structure. We identified rtn1Delta in a systematic screen for yeast mutants with altered ER morphology. The ER in rtn1Delta cells is predominantly cisternal rather than reticular, yet the net surface area of ER is not significantly changed. Rtn1-green fluorescent protein (GFP) associates with the reticular ER at the cell cortex and with the tubules that connect the cortical ER to the nuclear envelope, but not with the nuclear envelope itself. Rtn1p overexpression also results in an altered ER structure. Rtn proteins are found on the ER in a wide range of eukaryotes and are defined by two membrane-spanning domains flanking a conserved hydrophilic loop. Our results suggest that Rtn proteins may direct the formation of reticulated ER. We independently identified Rtn1p in a proteomic screen for proteins associated with the exocyst vesicle tethering complex. The conserved hydophilic loop of Rtn1p binds to the exocyst subunit Sec6p. Overexpression of this loop results in a modest accumulation of secretory vesicles, suggesting impaired exocyst function. The interaction of Rtn1p with the exocyst at the bud tip may trigger the formation of a cortical ER network in yeast buds.
Assuntos
Retículo Endoplasmático/ultraestrutura , Proteínas de Membrana/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/ultraestrutura , Retículo Endoplasmático/metabolismo , Deleção de Genes , Membranas Intracelulares/metabolismo , Proteínas de Membrana/análise , Proteínas de Membrana/genética , Proteínas de Membrana Transportadoras , Organelas/metabolismo , Organelas/ultraestrutura , Proteômica , Canais de Translocação SEC , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/análise , Proteínas de Saccharomyces cerevisiae/genética , Vesículas Secretórias/metabolismoRESUMO
The endoplasmic reticulum (ER) is a fundamental organelle required for protein assembly, lipid biosynthesis, and vesicular traffic, as well as calcium storage and the controlled release of calcium from the ER lumen into the cytosol. Membranes functionally linked to the ER by vesicle-mediated transport, such as the Golgi complex, endosomes, vacuoles-lysosomes, secretory vesicles, and the plasma membrane, originate largely from proteins and lipids synthesized in the ER. In this review we will discuss the structural organization of the ER and its inheritance.
Assuntos
Retículo Endoplasmático/fisiologia , Células Eucarióticas/fisiologia , Células Procarióticas/fisiologia , Animais , Retículo Endoplasmático/química , Complexo de Golgi , Humanos , Metabolismo dos Lipídeos , Organelas , Dobramento de ProteínaRESUMO
In Saccharomyces cerevisiae, the endoplasmic reticulum (ER) is found along the cell periphery (cortical ER) and nucleus (perinuclear ER). In this study, we characterize a novel ER protein called Ice2p that localizes to the cortical and perinuclear ER. Ice2p is predicted to be a type-III transmembrane protein. Cells carrying a genomic disruption of ICE2 display defects in the distribution of cortical ER in mother and daughter cells. Furthermore, fluorescence imaging of ice2delta cells reveals an abnormal cortical ER tubular network morphology in both the mother cell and the developing bud. Subcellular fractionation analysis using sucrose gradients corroborate the data from the fluorescence studies. Our findings indicate that Ice2p plays a role in forming and/or maintaining the cortical ER network in budding yeast.