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M Muñiz

Publications and source records attributed to M Muñiz.

11 recordsLinked to original sources
PubMed2001

Protein sorting upon exit from the endoplasmic reticulum.

It is currently thought that all secretory proteins travel together to the Golgi apparatus where they are sorted to different destinations. However, the specific requirements for transport of GPI-anchored proteins from the endoplasmic reticulum to the Golgi apparatus in yeast could be explained if protein sorting occurs earlier in the pathway. Using an in vitro assay that reconstitutes a single round of budding from the endoplasmic reticulum, we found that GPI-anchored proteins and other secretory proteins exit the endoplasmic reticulum in distinct vesicles. Therefore, GPI-anchored proteins are sorted from other proteins, in particular other plasma membrane proteins, at an early stage of the secretory pathway. These results have wide implications for the mechanism of protein exit from the endoplasmic reticulum.

Adenosine Triphosphatases↗
PubMed2000

The Emp24 complex recruits a specific cargo molecule into endoplasmic reticulum-derived vesicles.

Members of the yeast p24 family, including Emp24p and Erv25p, form a heteromeric complex required for the efficient transport of selected proteins from the endoplasmic reticulum (ER) to the Golgi apparatus. The specific functions and sites of action of this complex are unknown. We show that Emp24p is directly required for efficient packaging of a lumenal cargo protein, Gas1p, into ER-derived vesicles. Emp24p and Erv25p can be directly cross-linked to Gas1p in ER-derived vesicles. Gap1p, which was not affected by emp24 mutation, was not cross-linked. These results suggest that the Emp24 complex acts as a cargo receptor in vesicle biogenesis from the ER.

Antibodies↗
PubMed2000

Intracellular transport of GPI-anchored proteins.

In eukaryotic cells, a subset of proteins are attached to the external leaflet of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. There is substantial evidence suggesting that these GPI-anchored proteins are clustered in sphingolipid-sterol microdomains or rafts. Since the precursors of these microdomain components are synthesized mainly in the endoplasmic reticulum, it is possible that microdomain assembly occurs during transport along the exocytic route. A sorting mechanism for GPI-anchored proteins using sphingolipid microdomains as selective platforms for vesicle budding has been proposed to operate at different steps in the secretory pathway. Here, we discuss this sorting model in the context of the data obtained from different biological and artificial systems, in addition to other particularities of the intracellular transport of the GPI-anchored proteins.

Animals↗
PubMed2000

Variation in tomato host response to Bemisia tabaci (Hemiptera: Aleyrodidae) in relation to acyl sugar content and presence of the nematode and potato aphid resistance gene Mi.

Two commercial cultivars of tomato, Alta and Peto 95, the accession line number LA716 of Lycopersicon pennellii and lines 94GH-006 and 94GH-033 (backcrosses between Peto 95 and LA716), with different leaf acyl sugar contents were screened for resistance to Bemisia argentifolii Bellows & Perring (corresponding to the Spanish B-biotype of Bemisia tabaci (Gennadius)), in greenhouse- and field-no-choice experiments. There was no oviposition on LA716 (with the highest acyl sugar content) while the greatest fecundity and fertility values were observed on the cultivar Alta (no acyl sugar content). However, no clear relationship was found between the low acyl sugar content in the other tomato cultivars tested and whitefly reproduction. Thus, resistance to B. tabaci did not appear to correlate with acyl sugar content below a threshold level of 37.8 microg cm-2 leaf. In a greenhouse choice-assay, B. tabaci exhibited reduced host preference and reproduction on the commercial tomato cultivars Motelle, VFN8 and Ronita all of which carry the Mi gene resistance to Meloidogyne nematodes and the aphid Macrosiphum euphorbiae (Thomas), than on the Mi-lacking cultivars Moneymaker, Rio Fuego and Roma. When data of Mi-bearing plants were pooled, the mean values for daily infestation and pupal production of B. tabaci were significantly lower than those of Mi-lacking plants. This reflected a level of antixenosis- and antibiosis-based resistance in commercial tomato and indicated that Mi, or another closely linked gene, might be implicated in a partial resistance which was not associated either with the presence of glandular trichomes or their exudates. These findings support the general hypothesis for the existence of similarities among the resistance mechanisms to whiteflies, aphids and nematodes in commercial tomato plants.

Animals↗
PubMed1998

Diva, a Bcl-2 homologue that binds directly to Apaf-1 and induces BH3-independent cell death.

We have identified and characterized Diva, which is a novel regulator of apoptosis. Sequence analysis revealed that Diva is a member of the Bcl-2 family of proteins containing Bcl-2 homology domain 1, 2, 3, and 4 (BH1, BH2, BH3, and BH4) regions and a carboxyl-terminal hydrophobic domain. The expression of Diva mRNA was detected in multiple embryonic tissues but was restricted to the ovary and testis in adult mice. The expression of Diva promoted the death of 293T, Ramsey, and T47D cells as well as that of primary sensory neurons, indicating that Diva is a proapoptotic protein. Significantly, Diva lacks critical residues in the conserved BH3 region that mediate the interaction between BH3-containing proapoptotic Bcl-2 homologues and their prosurvival binding partners. Consistent with this, Diva did not bind to cellular Bcl-2 family members including Bcl-2, Bcl-XL, Bcl-w, Mcl-1, and A1/Bfl-1. Furthermore, mutants of Diva lacking the BH3 region fully retained their proapoptotic activity, confirming that Diva promotes apoptosis in a BH3-independent manner. Significantly, Diva interacted with a viral Bcl-2 homologue (vBcl-2) encoded by the Kaposi's sarcoma-associated herpesvirus. Consistent with these associations, apoptosis induced by Diva was inhibited by vBcl-2 but not by Bcl-XL. Importantly, Diva interacted with Apaf-1, an adapter molecule that activates caspase-9, a central death protease of the apoptotic pathway. The expression of Diva inhibited the binding of Bcl-XL to Apaf-1, as determined by immunoprecipitation assays. Thus, Diva represents a novel type of proapoptotic Bcl-2 homologue that promotes apoptosis independently of the BH3 region through direct binding to Apaf-1, thus preventing Bcl-XL from binding to the caspase-9 regulator Apaf-1.

Amino Acid Sequence↗
PubMed1998

Calphostin C induces selective disassembly of the Golgi complex by a protein kinase C-independent mechanism.

Intact cells incubated with calphostin C, an inhibitor of the regulatory domain of protein kinase C, showed fragmentation and dispersal of the Golgi complex by a light-dependent mechanism. At the ultrastructural level Golgi stacks were replaced by clusters of vesicles and short tubules that resembled the Golgi remnants present in control mitotic cells. Vesicle-mediated transport processes along both the exocytic and endocytic routes were also inhibited by calphostin C treatment. Golgi disassembly, however, was not due to protein kinase C inhibition since several inhibitors of the catalytic domain did not cause a similar effect. In contrast, pretreatment with phorbol 12-myristate 13-acetate partly protected the Golgi complex from disassembly by calphostin C. The in vitro effect was shown to be reversible, required both cytosol and ATP and it was inhibited by pretreatment of the Golgi membranes with trypsin but not with high salt. These results suggest the interaction of calphostin C with a structural Golgi protein containing a phorbol ester-binding domain and necessary for the stability of this organelle during interphase.

Adenosine Triphosphate↗
PubMed1998

[Prognostic value of the initial response of serum estradiol levels to stimulation with GnRH analogues in in vitro fertilization].

Studying the value prognostic of the answer of the Estradiol (E2) levels to the stimulation with agonists of GnRH (GnRH-a) during early follicular phase. Retrospective, analytic and comparative study. From January 1, 1993 to December 31, 1995 403 IVF cycles were studied. According to E2 levels after stimulation with GnRH-a four Patterns were assembled. Pattern A (n = 115), B (n = 22), C (n = 11), and D (n = 3). Those was included patients managed in short luteal phase in this study, (FLC) (n = 153) and long luteal phase (FLL) (n = 97) were groups control. The rate of oocyte retrieve was greater in the patterns, A, B, FLC and FLL with respect to the patron D (p = 0.001). The pregnancy rate was higher in A, B, FLC and FLL (25%, 19%, 22% and 25% respectively (p = 0.05). The age was old in C and D and luteal phase with an average of 35 +/- 2 (p = 0.001). The patterns had a better answer, represented by the increment of the estradios (E2) levels response to the GnRH-a action of leuprolide acetate during the early follicular phase they went A and B. We conclude that the early E2 response patterns to leuprolide acetate is the best early prognostic indicator of IVF outcome.

Adult↗
PubMed1997

Protein kinase A activity is required for the budding of constitutive transport vesicles from the trans-Golgi network.

We have examined the role played by protein kinase A (PKA) in vesicle-mediated protein transport from the trans-Golgi network (TGN) to the cell surface. In vivo this transport step was inhibited by inhibitors of PKA catalytic subunits (C-PKA) such as the compound known as H89 and a myristoylated form of the inhibitory peptide sequence contained in the thermostable PKA inhibitor. Inhibition by H89 occurred at an early stage during the transfer of vesicular stomatitis virus G glycoprotein from the TGN to the cell surface. Reversal from this inhibition correlated with a transient increase in the number of free coated vesicles in the Golgi area. Vesicle budding from the TGN was studied in vitro using vesicular stomatitis virus-infected, permeabilized cells. Addition to this assay of C-PKA stimulated vesicle release while it was suppressed by PKA inhibitory peptide, H89, and antibody against C-PKA. Furthermore, vesicle release was decreased when PKA-depleted cytosol was used and restored by addition of C-PKA. These results indicate a regulatory role for PKA activity in the production of constitutive transport vesicles from the TGN.

Animals↗
PubMed1996

A regulatory role for cAMP-dependent protein kinase in protein traffic along the exocytic route.

The influence of protein kinase A activity on transport of newly synthesized vesicular stomatitis virus G glycoprotein along the exocytic pathway was examined. Transport of vesicular stomatitis virus G glycoprotein to the cell surface was inhibited by N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89), a selective inhibitor of protein kinase A. This block occurred at the exit of the Golgi complex, whereas transport through the Golgi compartments or from the endoplasmic reticulum to the Golgi was decreased in the presence of H-89. As judged by immunofluorescence endoplasmic reticulum to Golgi transport was accelerated in cells incubated with activators of protein kinase A such as isobutylmethylxanthine (IBMX) or forskolin (FK). Treatment with IBMX and FK also increased transport from the trans-Golgi network to the cell surface. During incubation with IBMX and FK, the organization of the Golgi complex was altered showing intercisternae fusion and miscompartmentalization of resident proteins. These structural changes affected both the kinetics of acquisition of endoglycosidase H resistance and transport activities. These data support a differential regulatory role for protein kinase A in different transport steps along the exocytic pathway. In particular, transport from the trans-Golgi network to the cell surface was dependent on protein kinase A activity. In addition, the results suggest the involvement of this enzyme on the maintenance of the Golgi complex organization.

1-Methyl-3-isobutylxanthine↗
PubMed1995

Trimeric G proteins regulate the cytosol-induced redistribution of Golgi enzymes into the endoplasmic reticulum.

Streptolysin O-permeabilized cells incubated with a high concentration (5-10 mg/ml) of cytosolic proteins and ATP-generating system exhibit redistribution into the endoplasmic reticulum (ER) of Golgi integral proteins (mannosidase II, galactosyltransferase, TGN 38), detected by immunofluorescence. In addition, mannosidase II is detected in the ER of cells exposed to a high concentration of cytosolic proteins and processed for immunolectron microscopy by immunoperoxidase. The redistribution process requires ATP and is not affected by previous microtubule depolymerization. Ultrastructural observations indicate that Golgi disassembly occurs by budding of coated vesicles. This stage of the process is inhibited by GTP-gamma S, AIF(3-5), transducin beta gamma subunits, and mastoparan, indicating the involvement of trimeric G proteins. At a later stage, vesicles lose their coats and fuse with the ER. This part of the process does not occur in cells incubated at either 15 degrees C or 20 degrees C, or exposed to N-ethylmaleimide. In cells treated with either cholera or pertussis toxin Golgi redistribution into the ER shows a 50-fold lower requirement for cytosolic factors than in untreated cells. These data suggest a regulatory role for both alpha s and alpha i trimeric G proteins in the normal Golgi-ER retrograde transport taking place in intact cells.

Adenosine Triphosphate↗