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PubMed1998

Localization of the P1 protein of potato Y potyvirus in association with cytoplasmic inclusion bodies and in the cytoplasm of infected cells.

The N-terminal P1 proteinase of potato virus Y (ordinary strain group isolate PVY-O) was expressed in E. coli. Antiserum was raised against the expressed protein and used to detect the viral proteins in infected tobacco leaf tissue by Western blotting and by electron microscopy with immunogold labelling. In the immunogold localization studies P1 protein was detected in association with the cytoplasmic inclusion bodies characteristic of PVY infections and in the cytoplasm of the infected plant cells. No significant P1 antibody binding with other plant cell organelles, or with the cell wall and plasmodesmata, was detected by immunogold labelling.

Animals↗

PubMed1994

Amoebapores, a family of membranolytic peptides from cytoplasmic granules of Entamoeba histolytica: isolation, primary structure, and pore formation in bacterial cytoplasmic membranes.

Three peptides with pore-forming activity were isolated from the cytoplasmic granules of pathogenic Entamoeba histolytica by acidic extraction, gel filtration and reversed-phase high-performance liquid chromatography. Partial amino acid sequence analysis of the three active peptides revealed that the most abundant of them was amoebapore and the other two were isoforms thereof. Cloning and sequencing of genomic DNA resolved the amino acid sequence of the two newly recognized peptides. The three peptides designated amoebapores A, B and C were found to have the same molecular size but to differ markedly in their primary structure, although all six cysteine residues are conserved. Despite sequence divergence, structural implications predict for the three peptides a similar amphipathic alpha-helical conformation stabilized by disulphide bonds. All three isoforms exhibit pore-forming activity toward lipid vesicles, but they differ in their kinetics. They also are capable of perturbing the integrity of bacterial cytoplasmic membranes and thereby kill Gram-positive bacteria. The amoebapores represent a distinct family of membrane-active peptides that may function intracellularly as antimicrobial agents but may also confer cytolytic activity on the parasite.

Amino Acid Sequence↗

PubMed2000

Evidence for Gal3p's cytoplasmic location and Gal80p's dual cytoplasmic-nuclear location implicates new mechanisms for controlling Gal4p activity in Saccharomyces cerevisiae.

Genetics and in vitro studies have shown that the direct interaction between Gal3p and Gal80p plays a central role in galactose-dependent Gal4p-mediated GAL gene expression in the yeast Saccharomyces cerevisiae. Precisely how Gal3p-Gal80p interaction effects induction is not clear. It has been assumed that Gal3p interacts with Gal80p in the nucleus upon galactose addition to release Gal80p inhibition of Gal4p. Although Gal80p has been shown to possess nuclear localization signal (NLS) peptides, the subcellular distribution of neither Gal80p nor Gal3p was previously determined. Here we report that Gal3p is located in the cytoplasm and apparently excluded from the nucleus. We show that Gal80p is located in both the cytoplasm and the nucleus. Converting Gal80p into a nucleus-localized protein (NLS-Gal80p) by exogenous NLS addition impairs GAL gene induction. The impaired induction can be partially suppressed by targeting Gal3p to the nucleus (NLS-Gal3p). We document a very rapid association between NLS-Gal3p and Gal80p in vivo in response to galactose, illustrating that the nuclear import of Gal80p is very rapid and efficient. We also demonstrate that nucleus-localized NLS-Gal80p can move out of the nucleus and shuttle between nuclei in yeast heterokaryons. These results are the first indication that the subcellular distribution dynamics of the Gal3 and Gal80 proteins play a role in regulating Gal4p-mediated GAL gene expression in vivo.

Cell Nucleus↗

PubMed1984

Formation of cytoplasm-containing vesicles from double-walled coated invaginations containing oligodendrocytic cytoplasm at the axon-myelin sheath interface in adult mammalian central nervous system.

Double-walled coated invaginations ( DWCIs ) of the axolemma, containing oligodendroglial cytoplasm and presumably giving rise to double-walled coated vesicles ( DWCVs ), are observed in the central nervous system (CNS) of adult monkey and mouse. By means of serial sections, DWCIs are clearly distinguishable from the also observable adaxonal invaginations of oligodendroglial cytoplasm. DWCIs range in diameter from 60 to 190 nm with a circular or oval cross section. Their incidence in different regions of the CNS has been determined and evidence is adduced that DWCIs occur randomly along the axolemma.

Animals↗

PubMed1996

Ketoisocaproic acid and leucine increase cytoplasmic pH in mouse pancreatic B cells: role of cytoplasmic Ca2+ and pH-regulating exchangers.

The effects of nonglucose nutrient insulin secretagogues on cytoplasmic pH (pHi) in pancreatic B cells are unclear. These were studied with intact mouse islets loaded with BCECF and stimulated with ketoisocaproic acid (KIC) or leucine, which, unlike glucose, are exclusively metabolized in mitochondria. The changes in pHi were compared to those in cytoplasmic Ca2+ ([Ca2+]i; islets loaded with fura-2), metabolism [NAD(P)H fluorescence], and insulin release. In HCO3- buffer containing 3 mM glucose, 10 mM KIC produced a rapid and sustained increase in metabolism, [Ca2+]i, pHi, and insulin release. In HEPES buffer, the increase in metabolism, [Ca2+]i, and release were also rapid but not as sustained, whereas the alkalinization was delayed. The changes in release thus follow a time course more similar to that of [Ca2+]i and metabolism than to that of pHi. The role of [Ca2+]i in pHi changes was next examined. A similar rapid rise in pHi was produced by KIC in both HCO3- and HEPES buffers when its effects on [Ca2+]i were prevented, whether [Ca2+]i was kept low (4.8 mM KCl plus diazoxide) or high (30 mM KCl plus diazoxide). When the Na(+):H+ exchanger was blocked by dimethylamiloride, the alkalinizing effect of KIC was unaffected in HCO3- buffer, indicating that it does not result from an activation of this exchanger. In HEPES buffer, however, KIC strongly decreased pHi unless the rise in [Ca2+]i was prevented, in which case KIC increased pHi. When the HCO3-/Cl-exchanger was blocked by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), the effect of KIC in HCO3- buffer became similar to that in HEPES buffer without or with DIDS. The effects of leucine on pHi were similar to those of KIC. In conclusion, the effect of KIC and leucine on islet cell pHi, like that of glucose, is the complex result of an alkalinizing action of their metabolism and an acidifying action of the [Ca2+]i rise that they also produce. Compensation of this acidification is achieved by operation of the pHi-regulating exchangers, of which the HCO3-/Cl- exchanger plays a predominant role.

Animals↗

PubMed1982

Phenylalanyl-tRNA synthetases from hen liver cytoplasm and mitochondria, yeast cytoplasm and mitochondria, and from Escherichia coli: substrate specificity relationship with regard to ATP analogs.

Twelve structural analogs of ATP have been tested in the aminoacylation reaction of phenylalanyl-tRNA synthetases from hen liver cytoplasm and mitochondria, yeast cytoplasm and mitochondria and E. coli. Three compounds are substrates for all five phenylalanyl-tRNA synthetase, three are completely inactive, while the other ATP analogs show differing properties with the different enzymes. Their Km, Ki and V values have been determined. The importance of the amino group in Position 6, the nitrogen in Position 7 and an unsubstituted Position 8 of the purine moiety as well as the supposed anti-conformation of the glycosidic bond and coordination of the magnesium cation to N-7 appear to be conserved through evolution. Bulky substituents on the 2' and 3' of the ribose moiety are generally not tolerated. Graduation of substrate properties of some analogs are similar for the intracellular heterotopic isoenzymes from yeast and hen liver.

Adenosine Triphosphate↗

PubMed1999

Atypical Wegener's granulomatosis with positive cytoplasmic antineutrophil cytoplasmic antibodies, ophthalmologic manifestations, and slowly progressive renal failure without respiratory tract involvement.

A 68-year-old woman had microscopic hematuria and proteinuria since the age of 50. She also had hearing impairment, arthralgia, retinal embolism, peripheral arterial occlusion of the right foot and chronic renal failure during the course. At the age of 68, she had progressive renal failure and nephrotic syndrome with high titers of serum cytoplasmic antineutrophil cytoplasmic antibodies (c-ANCA). No evidence of respiratory tract involvement was found. Methylprednisolone pulse therapy and low dose cyclophosphamide therapy ameliorated the renal failure and reduced the serum c-ANCA level. She, however, died on July 19, 1998 due to pulmonary fungal and pneumocystis carinii infection.

Aged↗

PubMed1999

Bronchiectasis with myeloperoxidase antineutrophil cytoplasmic antibody and bactericidal/permeability-increasing protein antineutrophil cytoplasmic antibody.

A 56-year-old woman was hospitalized for recurrent hemoptysis. She had been suffering from bronchiectasis for 4 years. Pseudomonas aeruginosa was persistently detected in her sputum. Serum was positive for Myeloperoxidase antineutrophil cytoplasmic antibody (MPO-ANCA) and bactericidal/permeability-increasing protein antineutrophil cytoplasmic antibody (BPI-ANCA). She underwent lung resection. Histopathologically, the resected lung showed bronchiectasis with pulmonary fibrosis but did not show vasculitis. Her serum became negative for the ANCAs after the operation. To date, she has no recurrence of hemoptysis. We discuss this case of bronchiectasis with MPO-ANCA and BPI-ANCA and suggest a possible role for ANCAs in chronic airway infection.

Antibodies, Antineutrophil Cytoplasmic↗

PubMed2006

Cytoplasmic antineutrophil cytoplasmic antibody positive pauci-immune glomerulonephritis associated with infectious endocarditis.

Renal deterioration often occurs in cases of infectious endocarditis (IE), but, IE- associated nephritis with rapidly progressive glomerulonephritis (RPGN) is rare. Patients with severe infection (e.g., IE) sometimes show positivity for cytoplasmic antineutrophil cytoplasmic antibodies (C-ANCA). Therefore, diagnosis and treatment are very difficult in cases of RPGN with IE and positivity for C-ANCA. Such cases are rare, only 12 have been reported in the English literature. Herein, we describe the case of a 50-year-old man who presented with RPGN with IE and tested positively for C-ANCA. He was referred to our hospital because of leg edema, purpura and renal dysfunction. Laboratory tests revealed serum creatinine elevation and positivity for C-ANCA and proteinase 3-specific (PR3)-ANCA. RPGN and acute renal failure were diagnosed. Hemodialysis and steroid therapy were started. Streptococcus oralis was isolated by blood culture. Transthoracic echocardiography revealed grade III mitral valve insufficiency with two vegetations. Therefore, IE was diagnosed. The steroid therapy was stopped, and antibiotic therapy was begun. Because there was no improvement, surgical therapy was performed. The operation was successful, but the patient died of brain hemorrhage. Our experience in this case indicates C/PR3-ANCA positive RPGN must be ruled out in patients with infectious disease, particularly IE, together with renal symptoms, and renal biopsy should be performed.

Acute Kidney Injury↗

PubMed1992

"False positive" perinuclear and cytoplasmic anti-neutrophil cytoplasmic antibody results leading to misdiagnosis of Wegener's granulomatosis and/or microscopic polyarteritis.

The antineutrophil cytoplasmic antibody (ANCA) test has been shown to be important in helping to confirm the diagnosis and following the clinical course of microscopic polyarteritis and Wegener's granulomatosis. So called "false positive" test results have been reported, but usually in patients without any clinical evidence of these diseases, and the "false positive" result ignored. I wish to report 4 cases, in which a diagnosis of microscopic polyarteritis/Wegener's granulomatosis was considered as part of the differential diagnosis based on the clinical findings. The ANCA test was positive for cytoplasmic staining in 2 cases and perinuclear in 2 others. The combination of a positive ANCA result and the clinical possibility of Wegener's granulomatosis and/or microscopic polyarteritis resulted in the prescription of immunosuppressive treatment, with the consequent mortality of one patient and significant morbidity in two of the other cases.

Adult↗

PubMed2005

Rye SCAR markers for male fertility restoration in the P cytoplasm are also applicable to marker-assisted selection in the C cytoplasm.

The study aimed at testing the usefulness of recently developed SCAR markers on rye (Secale cereale L.) chromosome 4R in hybrid breeding based on the C source of male sterility-inducing cytoplasm. Of 10 markers studied, 4 revealed polymorphisms between 2 inbred lines (544cms-C and Ot0-20) crossed to develop F2 and BC1 mapping populations. Analyses performed on 94 F2 and 93 BC1 plants allowed to extend a formerly constructed genetic map of chromosome arm 4RL. Three SCAR markers (SCP14M55, SCP15M55 and SCP16M58) were mapped in the vicinity of gene Rfc1, which restores male fertility in the C cytoplasm. The 3 tested SCAR markers proved to be effective in marker-assisted selection (MAS) for male fertility/sterility.

Breeding↗

PubMed1985

[Use of cell enucleation in studying the stability of cytoplasmic organelles and cytoplasm organization].

A study was made of the viability and ultrastructure of cytoplasts produced by enucleation of cytochalasin-induced A9 cells in suspension. These cytoplasts are in general as viable as cells enucleated in the monolayer. The organization of the cytoplasm, i.e. a specific distribution of cytoplasmic organelles, is conserved for at least 24 hours in the absence of the nucleus. This fact may reflect a high degree of autonomy of the cytoskeleton.

Animals↗

PubMed1996

The association between human leukocyte antigens (HLA) and cytoplasmic-antineutrophil cytoplasmic antibody (cANCA)-positive Wegener's granulomatosis in a Japanese population.

The present study examined the association between various human leukocyte antigens (HLA) and cytoplasmic-antineutrophil cytoplasmic antibody (cANCA)-positive Wegener's granulomatosis (WG) in Japanese subjects to determine whether HLA antigens are involved in the pathogenesis of this disease. The study involved 16 subjects with cANCA-positive WG treated in our department. HLA-typing of the lymphocytes was performed using a lymphocyte microcytotoxicity assay. Of the subjects with cANCA-positive WG, 62.5% (10/16) were positive for HLA-DR9, as compared to 26% of the healthy control subjects. This HLA-DR9 elevation was statistically significant (p < 0.01, Pc < 0.05); we also noted a weaker association between HLA-B55 and cANCA-positive WG (p < 0.05). The results indicate that an association may exist between certain HLA-class allotypes and WG.

Antibodies, Antineutrophil Cytoplasmic↗

PubMed1996

[The homology of the primary structure of the third cytoplasmic domains in rhodopsin-type receptors and of the cytoplasmic tail in the beta-subunit of the insulin receptor].

In the last years evidence has been provided that along with the main mechanism of insulin signal transduction, that involves the ligand-induced tyrosine kinase cascade mediating the majority of biological effects of insulin, there is another signalling mechanism involving the heterotrimeric G-proteins. To detect the regions of amino acid sequences (AAS), responsible for interaction between the insulin receptor (IR) and G-proteins, a comparative analysis was carried out of the primary structure of third cytoplasmic domains (CD3) of rhodopsin-related receptors (RRR) determining RRR coupling to G-proteins, and of the primary structure of the cytoplasmic tail of IR beta-subunit. It is shown that AAS of juxtamembrane, ATP-binding and C-terminal domains of IR include long regions, homologous to the regions of RRR CD3, which are functionally important for coupling with G-proteins. AAS alignment of homologous RRR CD3 and beta-subunit of IR regions was made. It is found that the clusters of basic amino acids (e.g. forming BBXXB-motifs) and serine residues, located in RRR, coincide with similar amino acids present in IR primary structure. It is known that: 1) the BBXXB-motifs participate in the interaction with G-protein molecules, and 2) serine residues, the targets for phosphorylation by protein kinases, are included in regulation of hormonal signalling for both the receptor types. So, in the primary structure of IR beta-subunit, the regions were identified, which can be molecular determinants included in specific interaction with G-proteins of different types.

Amino Acid Sequence↗

PubMed1998

The presence of cytoplasmic antineutrophil cytoplasmic antibodies (C-ANCA) in the course of subacute bacterial endocarditis with glomerular involvement, coincidence or association?

Antineutrophil cytoplasmic antibodies positivity with cytoplasmic pattern (C-ANCA) and proteinase-3 (PR-3) specificity was found in two patients with both subacute bacterial endocarditis (SBE) and glomerular involvement. Renal biopsy showed membranoproliferative glomerulonephritis in one case and focal segmental glomerulonephritis in the second case. Immunofluorescence study showed granular immune deposits in both cases evocating immune complex glomerulonephritis. Renal and biological manifestations disappeared with clinical improvement secondary to antibiotherapy. Physicians have to consider the possible occurrence of such C-PR-3 ANCA, claimed to be specific markers for Wegener's granulomatosis, in infectious diseases such as SBE. Hence we focus on the necessity of performing a renal biopsy with light microscopy and immunofluorescence studies in all patients with ANCA associated glomerular disease.

Antibodies, Antineutrophil Cytoplasmic↗

PubMed2003

A cytoplasmic xylanase (XynX) of Aeromonas caviae ME-1 is released from the cytoplasm to the periplasm by osmotic downshock.

Aeromonas caviae ME-1 is a multiple xylanase-producing gram-negative bacterium which was isolated from the gut contents of a wild silkworm, Samia cynthia pryeri. One of the xylanases produced by A. caviae ME-1, XynX (38 kDa, family 10 xylanase), hydrolyzes xylan to xylobiose and xylotetraose as final degradation products. Generally, xylanases are extracellular or cell surface enzymes. However, XynX is not exported to the extracellular fluid by A. caviae ME-1 and an Escherichia coli transformant harboring the xynX gene. In this study, we investigated the intracellular localization of XynX in A. caviae ME-1 and an E. coli transformant. XynX was found in the cytoplasm when the cells were grown under normal culture conditions. However, XynX was released from the cytoplasm to the periplasm during osmotic downshock. This release of XynX in the E. coli transformant was blocked in the presence of gadolinium chloride, which has been reported to be an inhibitor of bacterial mechanosensitive channels.

Journal Article↗

PubMed1986

Fractionation of Rhizobium leguminosarum cells into outer membrane, cytoplasmic membrane, periplasmic, and cytoplasmic components.

Rhizobium leguminosarum cells were separated into four distinct fractions by using density gradient centrifugation for the separation of the outer and cytoplasmic membranes and lysozyme-EDTA treatment of whole cells for the isolation of the periplasmic and cytoplasmic fractions. These methods allowed the subcellular localization of R. leguminosarum proteins.

Bacterial Proteins↗

PubMedDate not supplied

Anti-neutrophil cytoplasm antibodies can recognize vascular endothelial cell-bound anti-neutrophil cytoplasm antibody-associated autoantigens.

Anti-neutrophil cytoplasm antibodies (ANCA) are strongly associated with the development of systemic vasculitis. Myeloperoxidase and proteinase-3 have been identified as targets for P-ANCA and C-ANCA, respectively. Both enzymes are released from neutrophil azurophil granules following neutrophil activation and both are highly cationic. Purified myeloperoxidase is demonstrated to bind non-convalently to endothelial cell membranes, to retain its enzymic function following binding, and to retain its antigenicity for P-ANCA. Endothelial cell-bound myeloperoxidase enhances complement-dependent cytotoxicity of some P-ANCA sere that also contain anti-endothelial cell antibodies. Studies using purified proteinase-3 show that it also can bind to endothelial cells and be recognized by C-ANCA. The interactions of myeloperoxidase and proteinase-3 with endothelial cells and ANCA may thus contribute to the development of vascular injury in patients with systemic vasculitis.

Antibodies, Antineutrophil Cytoplasmic↗