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A Pillez

Publications and source records attributed to A Pillez.

12 recordsLinked to original sources
PubMed1999

The transmembrane domain of hepatitis C virus glycoprotein E1 is a signal for static retention in the endoplasmic reticulum.

Hepatitis C virus (HCV) glycoproteins E1 and E2 assemble to form a noncovalent heterodimer which, in the cell, accumulates in the endoplasmic reticulum (ER). Contrary to what is observed for proteins with a KDEL or a KKXX ER-targeting signal, the ER localization of the HCV glycoprotein complex is due to a static retention in this compartment rather than to its retrieval from the cis-Golgi region. A static retention in the ER is also observed when E2 is expressed in the absence of E1 or for a chimeric protein containing the ectodomain of CD4 in fusion with the transmembrane domain (TMD) of E2. Although they do not exclude the presence of an intracellular localization signal in E1, these data do suggest that the TMD of E2 is an ER retention signal for HCV glycoprotein complex. In this study chimeric proteins containing the ectodomain of CD4 or CD8 fused to the C-terminal hydrophobic sequence of E1 were shown to be localized in the ER, indicating that the TMD of E1 is also a signal for ER localization. In addition, these chimeric proteins were not processed by Golgi enzymes, indicating that the TMD of E1 is responsible for true retention in the ER, without recycling through the Golgi apparatus. Together, these data suggest that at least two signals (TMDs of E1 and E2) are involved in ER retention of the HCV glycoprotein complex.

Amino Acid Sequence↗
PubMed1998

Hepatitis C virus glycoprotein complex localization in the endoplasmic reticulum involves a determinant for retention and not retrieval.

The hepatitis C virus (HCV) genome encodes two envelope glycoproteins (E1 and E2). These glycoproteins interact to form a noncovalent heterodimeric complex which in the cell accumulates in endoplasmic reticulum (ER)-like structures. The transmembrane domain of E2, at least, is involved in HCV glycoprotein complex localization in this compartment. In principle, ER localization of a protein can be the consequence of actual retention in this organelle or of retrieval from the Golgi. To determine which of these two mechanisms is responsible for HCV glycoprotein complex accumulation in the ER, the precise localization of these proteins was studied by immunofluorescence, and the processing of their glycans was analyzed. Immunolocalization of HCV glycoproteins after nocodazole treatment suggested an ER retention. In addition, HCV glycoprotein glycans were not modified by Golgi enzymes, indicating that the ER localization of these proteins is not because of their retrieval from the cis Golgi. Retention of HCV glycoprotein complexes in the ER without retrieval suggests that this compartment plays an important role for the acquisition of the envelope of HCV particles. A true retention in the ER was also observed for E2 expressed in the absence of E1 or for a chimeric protein containing the ectodomain of CD4 in fusion with the transmembrane domain of E2. These data indicate that, in HCV glycoprotein complex, the transmembrane domain of E2, at least, is responsible for true retention in the ER, without recycling through the Golgi.

Animals↗
PubMed1998

A retention signal necessary and sufficient for endoplasmic reticulum localization maps to the transmembrane domain of hepatitis C virus glycoprotein E2.

The hepatitis C virus (HCV) genome encodes two envelope glycoproteins (E1 and E2). These glycoproteins interact to formin a noncovalent heterodimeric complex which is retained in the endoplasmic reticulum (ER). To identify whether E1 and/or E2 contains an ER-targeting signal potentially involved in ER retention of the E1-E2 complex, these proteins were expressed alone and their intracellular localization was studied. Due to misfolding of E1 in the absence of E2, no conclusion on the localization of its native form could be drawn from the expression of E1 alone. E2 expressed in the absence of E1 was shown to be retained in the ER similarly to E1-E2 complex. Chimeric proteins in which E2 domains were exchanged with corresponding domains of a protein normally transported to the plasma membrane (CD4) were constructed to identify the sequence responsible for its ER retention. The transmembrane domain (TMD) of E2 (C-terminal 29 amino acids) was shown to be sufficient for retention of the ectodomain of CD4 in the ER compartment. Replacement of the E2 TMD by the anchor signal of CD4 or a glycosyl phosphatidylinositol (GPI) moiety led to its expression on the cell surface. In addition, replacement of the E2 TMD by the anchor signal of CD4 or a GPI moiety abolished the formation of E1-E2 complexes. Together, these results suggest that, besides having a role as a membrane anchor, the TMD of E2 is involved in both complex formation and intracellular localization.

Amino Acid Sequence↗
PubMed1997

Formation of native hepatitis C virus glycoprotein complexes.

The hepatitis C virus (HCV) glycoproteins (E1 and E2) interact to form a heterodimeric complex, which has been proposed as a functional subunit of the HCV virion envelope. As examined in cell culture transient-expression assays, the formation of properly folded, noncovalently associated E1E2 complexes is a slow and inefficient process. Due to lack of appropriate immunological reagents, it has been difficult to distinguish between glycoprotein molecules that undergo productive folding and assembly from those which follow a nonproductive pathway leading to misfolding and aggregation. Here we report the isolation and characterization of a conformation-sensitive E2-reactive monoclonal antibody (H2). The H2 monoclonal antibody selectively recognizes slowly maturing E1E2 heterodimers which are noncovalently linked, protease resistant, and no longer associated with the endoplasmic reticulum chaperone calnexin. This complex probably represents the native prebudding form of the HCV glycoprotein heterodimer. Besides providing a novel reagent for basic studies on HCV virion assembly and entry, this monoclonal antibody should be useful for optimizing production and isolation of native HCV glycoprotein complexes for serodiagnostic and vaccine applications.

Animals↗
PubMed1994

Immunocytochemical and in vitro autoradiographic evidence for a direct somatostatinergic modulation of the enkephalinergic hypothalamoseptal tract of the guinea-pig.

The present study was undertaken to determine whether the enkephalinergic hypothalamoseptal tract originating in the magnocellular dorsal nucleus in the guinea-pig brain is under the influence of somatostatin. In the first step, double immunocytochemical labeling of enkephalinergic cells and somatostatinergic fibers was combined at the light and electron microscopic levels in the magnocellular dorsal nucleus. As a second step, an in vitro radioautography was used to determine whether somatostatin receptors are present in the same area. A close relationship between somatostatin nerve endings and enkephalin perikarya was observed at both the light and electron microscopic levels. Contracts were more numerous in the ventral part of the magnocellular dorsal nucleus. Whenever synaptic images were clearly observable, they appeared symmetrical. In the same area, a moderate concentration of G-protein-coupled somatostatin binding sites was also visualized. These results suggest that somatostatin has a regulator role on the enkephalinergic hypothalamoseptal tract, directly at the level of the magnocellular dorsal nucleus.

Afferent Pathways↗
PubMed1993

Plasticity in expression of immunoreactivity for neuropeptide Y, enkephalins and neurotensin in the hypothalamic tubero-infundibular dopaminergic system during lactation in mice.

In lactating nursing vs lactating pup-deprived mice, single or multiple immunolabeling was performed to compare immunoreactivities (ir) for neuropeptide Y (NPY), enkephalins (ENK) and neurotensin (NT) in the tyrosine hydroxylase (TH)-ir (ENK) and neurotensin (NT) in the tyrosine hydroxylase (TH)-ir hypothalamic tubero-infundibular dopaminergic (TIDA) system. NPY-, ENK- and NT-irs were intensely expressed and coexisted in virtually all TH-ir endings in the median eminence (ME) of nursing mice. Removal of the pups induced a marked depletion of the peptide-irs from the ME TH-ir endings. In the arcuate nucleus (ARC) of colchicine-treated nursing mice which received peripheral injections of Fluoro-Gold (FG) to retrogradely label neuroendocrine cells, virtually all dorsal A12 TH-ir perikarya simultaneously expressed, with individual variations, NPY-, ENK- and NT-irs, and all contained FG. These results suggest that the synthesis of NPY, ENK and NT is enhanced in TIDA neurons during lactation and that these neuromessengers may be co-released together with DA from the ME to regulate the suckling-induced prolactin secretion at the hypothalamic and/or pituitary levels.

Animals↗
PubMed1984

[Immunohistochemical arguments in favor of co-localization of neuropeptides in hypothalamo-infundibular neuron systems].

Light and electron microscope methods which can be used to examine the possibilities of co-occurrence of peptides are reviewed. The results obtained using some of them are exposed and discussed, concerning i/the light microscope demonstration of the CRF/vasopressin coexistence in rat and guinea-pig. ii/the electron microscope demonstration of the granular colocalization of LH-RH/C-terminal ACTH and somatostatin/enkephalin in the guinea-pig median eminence.

Adrenalectomy↗
PubMed1983

Serotonin axons of the ependyma and circumventricular organs in the forebrain of the guinea pig. An immunohistochemical study.

The distribution of supraependymal nerve fibers (SEF) containing serotonin (5-HT) was investigated immunohistochemically in the forebrain of the guinea pig. The highest densities of immunoreactive axons were found in the pars centralis and the inferior horn of the lateral ventricle and also in the superior part of the third ventricle. Because of the special development of the choroid plexus in these ventricular regions, it is suggested that 5-HT SEF might be involved in the regulation of the composition of the cerebrospinal fluid. The ependyma lining the circumventricular organs located in the forebrain, was not observed to receive a significant 5-HT-SEF innervation. In the pituitary gland, a loose but constant network of 5-HT axons, resembling those which course in the anterobasal hypothalamus, arcuate nucleus and internal layer of the median eminence, was observed in the neural lobe. In the epiphysis, immunoreactive 5-HT was detected in all pinealocytes (the entire cell was filled with reaction product) and in fibers running between them.

Animals↗
PubMed1983

Ability of the CRF immunoreactive neurons of the paraventricular nucleus to produce a vasopressin-like material. Immunohistochemical demonstration in adrenalectomized guinea pigs and rats.

In the paraventricular nucleus of normal or adrenalectomized colchicine-treated guinea pigs and rats, corticotropin-releasing factor (CRF), vasopressin (VP) and oxytocin (OX) immunoreactivities were compared. In the control animals, respective stainings for these three peptides are distinct. Adrenalectomy resulted in the appearance of a VP-like staining in most of the CRF-immunoreactive neurons whereas OX staining remained distinct. It is suggested that the CRF/VP coexistence reflects the synergistic role of the two peptides.

Adrenalectomy↗
PubMed1983

[Presence of a substance immunologically related to somaliberin extracted from human pancreatic tumor (hpGRF) in neurons of the hypophysiotropic area in the guinea-pig and rat].

An antiserum raised against the synthetic human pancreatic tumour GRF (hpGRF) made it possible to localize an immunoreactive material in 100 nm granules of nerve terminals distributed in the external layer of the median eminence in Guinea-Pig and Rat. Moreover, especially in colchicine-treated Guinea-Pigs, numerous stained perikarya were observed, exclusively located in the arcuate nucleus. These results suggest that hypothalamic GRF is very similar, if not identical, to hpGRF.

Animals↗