Showing papers on "Cellular compartment published in 1983"

Hepatoma secretory proteins migrate from rough endoplasmic reticulum to Golgi at characteristic rates

Abstract: In eukaryotic cells, secretory proteins and glycoproteins migrate from the rough endoplasmic reticulum, their site of synthesis, through Golgi vesicles before being released from the cell. Cellular and viral integral plasma membrane glycoproteins are co-translationally inserted into the rough endoplasmic reticulum membrane and follow a similar pathway to the cell surface. Previous studies using endoglycosidase H (Endo H) suggested that in rat hepatoma cells the vesicular stomatitis virus (VSV) G protein, albumin and transferrin migrate from the rough endoplasmic reticulum to the Golgi apparatus at different rates. Here we show directly that in human hepatoma HepG2 cells, five secreted proteins mature from the rough endoplasmic reticulum to Golgi vesicles at characteristic rates which differ at least threefold. The results are incompatible with bulk-phase movement of the luminal contents of the endoplasmic reticulum, and suggest that there is a membrane-bound receptor that selectively mediates the transport of secretory proteins from the rough endoplasmic reticulum to the Golgi.

Alkaline phosphatase biosynthesis in the endoplasmic reticulum and its transport through the Golgi apparatus to the plasma membrane: cytochemical evidence.

Abstract: Enzyme induction of HeLa cell placental alkaline phosphatase with various agents such as prednisolone, sodium butyrate, hyperosmolality (NaCl), or combination of these inducers resulted in the appearance of enzyme activity in the rough endoplasmic reticulum, nuclear envelope, Golgi apparatus, and plasma membrane. In the Golgi apparatus, intense reaction product deposits tended to be concentrated on its trans side, with small vesicles and granules also being positively stained. Inhibition of protein synthesis with cycloheximide was followed by the disappearance of enzyme activity from these cytoplasmic organelles but not from the plasma membrane. Treatment with monensin, a secretory protein transport inhibitor, uniformly increased activity in the rough endoplasmic reticulum while causing marked dilatation of the intensely positive Golgi cisternae. These results suggest that intracellular alkaline phosphatase is newly synthesized in the endoplasmic reticulum and then passes en route through the Golgi appara...

Mécanismes d'action des hormones thyroïdiennes au niveau cellulaire

Abstract: Thyroid hormones exert pleiotropic effects at the cell level and several theories to define the initial site of action have been proposed. First mitochondria, nuclei or plasma membrane were considered as the main regulatory step. More recently, evidence of T3 and/or T4 binding sites present in different cellular compartments suggests the existence of more than one initial site of action. Binding sites from cytosol or plasma membrane and endoplasmic reticulum seem to participate either in the storage of thyroid hormones, or in their intracellular transport and metabolism. Mitochondrial and certain plasma membrane binding sites could be responsible for rapid effects of thyroid hormones at their level. Nuclear T3 binding sites located in the chromatin are considered as a true hormone receptor and are more documented in a structural and functional point of view. They may mediate thyroid hormone effects on the regulation of DNA transcription and the synthesis of active proteins. Their mechanism of action is nevertheless not well defined.

Transport and cleavage of bacterial pre-β-lactamase by mammalian microsomes

Abstract: Eucaryotic endoplasmic reticulum and bacterial inner cell membranes use strikingly similar mechanisms to transport proteins from one cellular compartment to another. Using a mammalian linked transcription-translation system, we show here that canine pancreatic microsomes accurately cleave off the signal sequence of β-lactamase, a secreted bacterial protein. We suggest that the apparent differences between bacterial and eukaryotic protein transport may involve only minor modulations of a profoundly conserved mechanism.

Subcellular compartmentalization of the progesterone receptor in cat uteri following the acute administration of progesterone.

Abstract: cytosol and nuclear progesterone receptors in the cat uterus were measured by Scatchard analysis to determine the relationships between dose of progesterone administered and the time following administration with the content of receptor in these two cellular compartments. Cats were ovariectomized, treated for 7 days with estradiol and then injected via the saphenous vein with progesterone. One uterine horn was removed prior to, and the other uterine horn after the injection of progesterone. The amount of cytosol receptor translocated was found to be dosedependent over the range of 0-200 pg of progesterone. A maximum of 40% of the cytosol receptor was depleted even when the amount of progesterone injected was increased. In non-estradiolprimed animals it was also found that approximately 40% of the cytosol receptor was depleted following a progesterone injection. Within 3 h of the injection of 300 pg of progesterone, the cytosol and nuclear receptor levels had returned to preinjection values. A second administration of progesterone at 1 or 3 h after the first injection of progesterone caused a partial depletion of the cytosol receptor and an increase in nuclear progesterone receptor concentration. These data suggest that the translocation of cytosol receptor and the appearance of nuclear receptor is dosedependent until approximately 40% of the cytosol receptor is depleted following a single injection of progesterone, that the retention of nuclear receptor after an acute injection of progesterone is of short duration (<1 h), and that the replenishment of cytosol receptor is complete within 3 h.

Alkaline Phosphatase Biosynthesis in the Endoplasmic Reticulum and Its Transport Through the Golgi Apparatus to the Plasma Membrane

Abstract: Enzyme induction of HeLa cell placental alkaline phosphatase with various agents such as prednisolone, sodium butyrate, hyperosmolality (NaCl), or combination of these inducers resulted in the appearance of enzyme activity in the rough endoplasmic reticulum, nuclear envelope, Golgi apparatus, and plasma membrane. In the Golgi apparatus, intense reaction product deposits tended to be concentrated on its trans side, with small vesicles and granules also being positively stained. Inhibition of protein synthesis with cycloheximide was followed by the disappearance of enzyme activity from these cytoplasmic organelles but not from the plasma membrane. Treatment with monensin, a secretory protein transport inhibitor, uniformly increased activity in the rough endoplasmic reticulum while causing marked dilatation of the intensely positive Golgi cisternae. These results suggest that intracellular alkaline phosphatase is newly synthesized in the endoplasmic reticulum and then passes en route through the Golgi apparatus to the plasma membrane. Accordingly, the present system could represent the biosynthesis, transport, and incorporation of the model cell surface enzyme protein to add to the vesicular stomatitus virus glyco-1 (VSV-G) protein and acetylcholine receptor model systems for studying the dynamics of cell surface protein genesis, transport, and membrane integration.

Effects of adrenaline on a compartment of slowly-exchangeable calcium in the perfused rat heart

Abstract: The effects of adrenaline on kinetically-distinct compartments of exchangeable calcium in the spontaneously-beating perfused rat heart were investigated under steady-state conditions using a 45Ca2+ outflow technique together with a nonlinear least-squares curve fitting procedure and 51Cr-EDTA to monitor the loss of freely-diffusible Ca2+ from the extracellular space. In addition to Ca2+ distributed in the vascular (compartment 1) and interstitial (compartment 2) spaces, the minimum number of kinetically-distinct compartments of cellular exchangeable Ca2+ (which include Ca2+ bound to extracellular sites on the sarcolemma) required to fit the data (compartments 3,4 and 5) was three. A system in which cellular compartment 3 is linked to the vascular space and compartments 4 and 5 are linked to the interstitial space was consistent with the data. For hearts perfused under control conditions, the fractional transfer rates for Ca2+ outflow from cellular compartments 3, 4 and 5 were 0.70, 0.11 and 0.017 min−1, respectively. Adrenaline increased the quantity of exchangeable Ca2+ in compartment 5 and decreased that present in the compartment 4. Evidence that compartment 5 includes mitochondrial exchangeable Ca2+ is discussed. It is concluded that one of the actions of adrenaline on the heart is to increase the quantity of exchangeable Ca2+ in the mitochondria.

Interactions between the rough endoplasmic reticulum and the nuclear membrane during increased proliferation of the endoplasmic reticulum.

Abstract: Flickinger (1978) has shown conclusively that when there is demand for membrane by the nucleus, such as during repair of a damaged nuclear envelope, several membrane continuities develop between the RER and the nuclear envelope. This experiment was set up to determine if reversal of flow of membrane from the nuclear envelope to the RER would occur during phenobarbital-stimulated increased synthesis of RER. During the increased synthesis of RER following the stimulation of liver cells, it was observed that there was increase in the number of membrane continuities between the RER and the nuclear envelope. Increased blebbing and vesicle formation from the nuclear envelope was also seen. These vesicles were liberated as RER into the cytoplasm. It is therefore suggested that the nuclear envelope and the RER are in a state of equilibrium (RER in equilibrium NE) and membrane can be transferred either way depending on demand through the many membrane continuities that develop between the two organelles.