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At least 145 records · Page 8Linked to original sources
PubMed1997

Crystal structure of cytoplasmic Escherichia coli peptidyl-prolyl isomerase: evidence for decreased mobility of loops upon complexation.

The structure of the unliganded form of the Escherichia coli cytoplasmic peptidyl-prolyl isomerase (ppiB gene product) in a new crystal form was determined by the molecular replacement method and refined to an R-factor of 16.1% at 2.1 A resolution. The enzyme crystallized in the orthorhombic C2221 space group with unit cell dimensions of a=44.7 A, b=68.2 A and c=102.0 A. Comparison with the reported structure of the enzyme complexed with the tripeptide substrate succinyl-Ala-Pro-Ala-p-nitroanilide revealed subtle changes that occur upon complex formation. There is evidence to suggest that two surface loops have significantly reduced mobility in the complexed structure.

Amino Acid Isomerases↗
PubMed1988

Wheat germ cytoplasmic ribosomes. Structure of ribosomal subunits and localization of N6,N6-dimethyladenosine by immunoelectron microscopy.

Cytoplasmic ribosomes have been isolated from wheat germ, and the structure of ribosomal subunits has been examined by electron microscopy of negatively stained preparations. Small (40 S) subunits show structural features generally regarded as characteristic of eukaryotic particles, while large (60 S) subunits show shapes that are equally well described by models of prokaryotic 50 S particles. Small subunit 18 S RNA contains 2 residues of N6,N6-dimethyladenosine 19 and 20 residues from the 3'-end (Hagenbüchle, O., Santer, M., Steitz, J. A., and Mans, R. J. (1978) Cell 13, 551-563). Nucleoside analysis by high performance liquid chromatography shows no other residues of this component in the RNA. Anti-dimethyladenosine immunoglobulins were reacted with wheat germ 40 S subunits, and the resulting complexes were studied by electron microscopy in order to localize the nucleoside. In about 90% of the complexes observed, antibody-subunit contact was consistent with a single binding site. We place the dimethyladenosine residues at or near the end of the platform of the 40 S particle in a position nearly equivalent to that previously identified in prokaryotic and chloroplast subunits (Trempe, M. R., and Glitz, D. G. (1981) J. Biol. Chem. 256, 11873-11879).

Adenosine↗
PubMed1975

Adaptation of the mitochondrial systems of Rhodotorula gracilis to low oxygen pressure.

The obligate aerobic yeast, Rhodotorula gracilis, was grown in a liquid minimal medium at 1 mm Hg partial pressure of oxygen, conditions under which growth (measured as the increase in total protein) is reduced to 30% of the maximum rate. A significant increase in the ratio between mitochondrial oxidative enzymes and total protein occurs rapidly under these conditions. A concurrent increase in the ratio area of mitochondria/area of cytoplasm was also observed. The relative increase in mitochondrial enzymes, oxidase activities and mitochondrial membranes is due to the inhibition affecting the increase in cytoplasmic structures more significantly than the increase in mitochondrial structures at low pO2. The difference between mitochondrial and cytoplasmic syntheses cannot be ascribed to changes in the availability of ATP but it might rest with some other oxygen-utilising process (pyrimidine redox coenzymes, synthesis of sterols). The experimental conditions studied appear to offer a valuable tool for the investigation of the relationships between mitochondrial and cytoplasmic structures.

Aerobiosis↗
PubMed2002

XpsG, the major pseudopilin in Xanthomonas campestris pv. campestris, forms a pilus-like structure between cytoplasmic and outer membranes.

GspG, -H, -I, -J and -K proteins are members of the pseudopilin family. They are the components required for the type II secretion pathway, which translocates proteins across the outer membrane of Gram-negative bacteria to the extracellular milieu. They were predicted to form a pilus-like structure, and this has been shown for PulG of Klebsiella oxytoca by using electron microscopy. In the present study, we performed biochemical analyses of the XpsG protein of Xanthomonas campestris pv. campestris and observed that it is a pillar-like structure spanning the cytoplasmic and outer membranes. Subcellular fractionation revealed a soluble form (SF) of XpsG, in addition to the membrane form. Chromatographic analysis of SF XpsG in the absence of a detergent indicated that it is part of a large complex (>440 kDa). In vitro studies indicated that XpsG is prone to aggregate in the absence of a detergent. We isolated and characterized a non-functional mutant defective in forming the large complex. It did not interfere with the function of wild-type XpsG and was not detectable in the SF. Moreover, unlike wild-type XpsG, which was distributed in both the cytoplasmic and outer membranes, it appeared only in the cytoplasmic membrane. When wild-type XpsG was co-expressed with His6-tagged XpsH but not with untagged XpsH, SF XpsG bound to nickel and co-eluted with XpsH. This result suggests the presence of other pseudopilin components in the XpsG-containing large-sized molecules.

Bacterial Proteins↗
PubMedDate not supplied

[The identification of internal symmetry in primary structure of cytoplasmic domains of human insulin receptor beta-subunit].

The graphic method of analysis for identification of internal symmetry centres in amino acid sequences (AAS) of insulin receptor (IR) cytoplasmic 'tail' was used. The method was based on the comparison of normal and conversional sequences (AAS or sequences of amino acid codon roots). It was shown, that the regions of IR cytoplasmic domains most important for receptor functional activity possess symmetrical structure. The regions participate in the autophosphorylation, tyrosine kinase activity, ATP-binding, interaction with different types of G-proteins, processing of receptor molecule, etc. The functionally important amino acids and their clusters either were included in the formation of the internal symmetry centres or were localized close to them.

Adenosine Triphosphate↗
PubMed1989

Intermediate filaments in biology and disease.

Intermediate filaments comprise a large heterogenous family of proteins in animal cells. Distinct from microfilaments and microtubules, they are a major component of the cytoskeleton and nuclear envelope. The expression of intermediate filament protein types is developmentally regulated and relatively cell type specific. Although there are at least five distinct classes of intermediate filament types, all the subunit proteins have similar structural features and appear to have envolved from some common gene ancestor. The cytoplasmic intermediate filaments exhibit a complex organization, forming associations with components of the nucleus, plasma membrane, and potentially other cytoplasmic structures such as microtubules. The specific function of this prominent cytoplasmic structure is currently unknown. However, the organization or expression of intermediate filaments is known to be altered in association with a variety of human diseases. Currently, specific antibodies to individual intermediate filament proteins are being used as an immunohistochemical aid for tumor typing in diagnostic pathology.

Animals↗
PubMed1976

Hatching glands in salmonid fishes, Salmo gairdneri, Salmo trutta, Salvelinus fontinalis and Salvelinus pluvius.

The unicellular hatching glands (UHGs) of four species of salmonid fish, Salmo gairdneri, Salmo trutta, Salvelinus fontinalis and Salvelinus pluvius were studied by light and electron microscopy. The UHGs are distributed on the epidermis of head and yolk sac, and on the epithelium of the mouth and gills. Since these cells are large and include dilated cisternae of the endoplasmic reticulum from the primitive to the mature stages they are conspicuous. Around the Golgi complexes, there are consecutive figures showing synthesis of secretory granules with close relationships to the Golgi vesicles. The secretory granules grow in size and vary in density during maturation; some have enclosed cytoplasmic structures. At the hatching stage, they are discharged with some cytoplasmic structures from the UHG which is located in the superficial layer of epithelium. After exhaustion of the secretory granules, the remainder of the contents of UHGs is liberated.

Animals↗
PubMed1997

The human p167 gene encodes a unique structural protein that contains centrosomin A homology and associates with a multicomponent complex.

The characterization of novel cytoplasmic, structural, and enzymatic proteins has been enhanced by a panel of monoclonal antibodies specific for protein substrates of transforming and nontransforming c-Src mutants. These protein substrates have included the focal adhesion kinase (FAK), cortactin, AFAP-110, p120CAS, and p130CAS. The monoclonal antibody 4G8 was generated as part of this panel of antibodies and was used to isolate the human gene for a 167-kD polypeptide. The cDNA sequence is 5,238 nucleotides in length with a predicted open reading frame consisting of 1,382 amino acids. The polypeptide is largely hydrophilic and highly charged. The central region of p167 has 88% identity with the entire 278-amino-acid encoded sequence of the murine centrosomin A gene. The carboxyl third of p167 contains a unique cluster of 10 amino acid repeats with the consensus sequence (A/M)DDDRGPRRG. The p167 protein was found primarily in the cytoplasm of lymphocytes and is part of a multicomponent protein complex with prominent members of 167, 120, 64, 45, 40, 38, and 25 kD. Finally, we illustrate the conservation of p167 and its associated complex, and demonstrate its expression in different human tissues and cell types. The data suggest that p167 is novel and has an important cellular function as a cytoplasmic structural protein.

Amino Acid Sequence↗
PubMed1960

Relation between basophilia and fine structure of cytoplasm in the fungus Allomyces macrogynus Em.

In a fungus, Allomyces macrogynus Em., staining tests have revealed changes in the location of cytoplasmic basophilia following different phases of the developmental cycle. These variations in location were used to observe which fine structures correspond to basophile and non-basophile areas of the cytoplasm. Hyphae, gametangia, zygotes, and plants were fixed at various developmental stages in OsO(4), pH 6.1, and embedded in vestopal. Sections were examined in the electron microscope. Comparison of basophile and non-basophile cytoplasms leads to the conclusion that cytoplasmic particles of 150 to 200 A in diameter are responsible for basophilia. The possibility of these particles being ribosomes is discussed and confirmed. The present paper also describes some observations on the fine structure of other cellular components of this fungus, such as nuclei, mitochondria, various granules, and flagella.

Allomyces↗
PubMed1979

Morphology of rabbit collecting duct.

Recently the assumed structural and functional homogeneity of the collecting duct (CD) has been questioned. The objective of this study was to determine if heterogeneity occurs in luminal surface membrane structure or in cytoplasmic configuration of cells in the collecting duct or both. Straight segments of cortical and medullary CD were examined in perfusion-fixed rabbit kidneys with scanning electron microscopy (SEM), light (LM) and transmission electron microscopy (TEM). Principal cells were the most abundant cells in all CD regions; intercalated cells comprised 37% of the cell population on the cortex, 18% in the outer medulla, and less than 1% in the inner medulla. SEM revealed two surface patterns among the ciliated principal cells: 1, located in the cortex and outer medulla, with few surface microvilli, and 2, located in the inner medulla, with abundant microvilli. Intercalated cells exhibited four distinctive luminal surface configurations: I, numerous short microvilli; II, both short and elongate microvilli; III, microplicae alone; and IV, both microvilli and microplicae. Intercalated cells with patterns I and II were predominant in the cortex, while cells with patterns III and IV were most common at the corticomedullary junction. TEM confirmed that marked variation existed in cytoplasmic structures of both principal and intercalated cells. These findings may either indicate the presence of several specific types of principal and intercalated cells or reflect different functional states of the principal and intercalated cells. Regardless of their significance, their presence must be considered in studies seeking to establish precise structural-functional relationships in this region of the rabbit renal tubule.

Animals↗
PubMed1998

Complexity of trypanosomatid endocytosis pathways revealed by Rab4 and Rab5 isoforms in Trypanosoma brucei.

Small G proteins of the Rab family are responsible for vesicle fusion and control flux during intracellular transport. Rab5 is important in endosome maturation and Rab4 in recycling of endocytic material. Three Rab5 isoforms identified so far in mammals and three in the yeast genome suggest that conservation of multiple Rab5 isoforms is required for sophisticated regulation of endocytosis. Trypanosoma brucei homologues of Rab5 and Rab4 (TbRab5A and TbRab4) have been identified. Here we report cloning of a second Rab5 homologue, TbRab5Bp. The TbRAB5A and -5B genes are not linked in the genome, and phylogenetic reconstruction indicates that multiple Rab5 isoforms in yeast, mammals, and trypanosomes evolved independently. Northern blots demonstrate that TbRab5A, -5B, and TbRab4 messages are expressed in bloodstream form (BSF) and procyclic forms of the parasite even though endocytosis is not very active in the latter form. mRNA levels of TbRab5A and -4 are constitutive. Multiple-sized TbRab5B messages at very low abundance are detected, with greater expression in BSF. Also, the TbRab5B mRNA has a large 3'-untranslated region suggestive of potentially complex regulation, and therefore TbRab5Bp may be an important regulator of differential endocytosis levels between BSF and procyclic stage parasites. Affinity purified antibodies raised to C-terminal peptide sequences of all three TbRab proteins recognized small vesicular cytoplasmic structures, which for TbRab5Ap and -5Bp are predominantly near the flagellar pocket. TbRab5Bp colocalizes with invariant surface glycoprotein 100 (ISG100), a protein entering the endocytotic pathway in BSF parasites, whereas in procyclic cells populations of vesicles stained with both TbRab5Ap and -5Bp substantially overlap; TbRab5 proteins are therefore components of the endocytotic pathway. TbRab4p localizes to vesicular structures throughout the cytoplasm, with some overlap with TbRab5Bp, but the majority occupying a different compartment to the TbRab5s. Therefore the trypanosome endosomal system has been functionally dissected for the first time; these reagents provide a unique opportunity for manipulation of the protozoan endosomal system to further our understanding of drug uptake mechanisms and virulence.

Amino Acid Sequence↗
PubMed1993

[Spontaneous and induced permeability of the tight junctions in the bladder epithelium of the frog Rana temporaria].

The spontaneous and induced alterations in paracellular permeability of the isolated frog urinary bladder were studied. For estimation of the tight junction (TJ) permeability, a fluorescent dye uranine was used. No morphological or functional evidences for the TJ opening were found in the autumn-winter period, both in the control and after the osmotic flow stimulation by 25 mg/ml pituitrine (P) or serosal mannitol (200 mM). In the spring, a spontaneous uranine efflux through the bladder wall from the mucosal solution was found in parallel with an increase in the basal water flow and a decrease in the responses of the bladder to P and serosal hypertonicity. At the same time, the intercellular substance remained to be a barrier for water. In the autumn-winter the TJ opening could be induced by adding sulfhydryl reagents, p-chloromercury-phenyl-sulfonic acid (PCMPS) or N-ethylmaleimide (NEM), to the serosal solution. However, NEM was effective only when added to the mucosal solution. The uranine efflux induced by sulfhydryl reagents was accompanied with a marked hydroosmotic flow in parallel with a diminished sensitivity to P, the barrier properties of intercellular substances were lost. As evidenced by electron microscopy, PCMPS induced a gradual destruction of TJ structures, while certain cytoplasmic structures were not affected. The uranine efflux in the autumn-winter frogs could be induced by an artificial osmotic gradient after preincubation of the bladders in the isotonic solution with P. Some possible intracellular mechanisms causing both spontaneous and induced alterations in the TJ permeability are discussed.

Animals↗
PubMed1983

Membrane-bound intranuclear inclusions in the Leydig cell of the Chinese hamster (Cricetulus griseus).

Membrane-bound intranuclear inclusions have been described, for the first time, in the Leydig cell of the Chinese hamster (Cricetulus griseus). The inclusions were not found in the 1-day-old animal, rarely found prior to sexual maturity, and commonly found in the sexually mature animals. The incidence of inclusions increases with aging. Their size and content varies greatly. They are surrounded by a single membrane and completely enclosed by nucleoplasm. Their close association with nuclear invaginations of cytoplasmic material, and their content of cytoplasmic structures along with some exhibiting the presence of trimetaphosphatase reaction product, suggest a cytoplasmic origin. This phenomenon involves the migration of cytoplasmic structures into the nucleus followed by detachment on the nucleoplasmic side. The presence of the inclusions is not an indication of an abnormality of the Leydig cell. The Leydig cell of the Chinese hamster may be an excellent model to study factors that initiate inclusion formation, and to determine the functional role of membrane-bound intranuclear inclusions.

Acid Anhydride Hydrolases↗
PubMed1991

Functional analysis of the tobacco mosaic virus tRNA-like structure in cytoplasmic gene regulation.

The 3'-untranslated region (UTR) of tobacco mosaic virus (TMV), which terminates in a tRNA-like structure, functionally substitutes for a poly(A) tail in both plant and animal cells. The addition of the TMV 3'-UTR to chimeric mRNA constructs increases their expression up to 100-fold, increasing both translational efficiency and mRNA stability. The domain largely responsible for the regulation maps to a 72 base region immediately upstream of the tRNA-like structure, however, the 3'-terminal, tRNA-like structure is required for full function. Its contribution is lost if separated from the upstream pseudoknot domain by as few as 5 bases or if 6 bases are removed from the 3'-terminus. Sequence addition to the 3'-terminus of the TMV 3'UTR or the upstream pseudoknot domain inhibits function in both tobacco and Chinese hamster ovary cells.

Animals↗
PubMed2001

A mechanism of intracellular timing and its cooperation with extracellular signals in controlling cell proliferation and differentiation, an amended hypothesis.

Various observations suggest that an intracellular timer is involved in the control of cell proliferation and differentiation that supplements control by extracellular signaling and depends on quantitative relations between cytoplasm and nucleus. To further elucidate the mechanism of this timer, we examined the results of experiments with mice in which cell cycle regulating genes were inactivated: the inactivation of negative cell cycle regulators extends cell proliferation, whereas inactivation of positive regulators decreases cell proliferation. We conclude that this is caused in the former case by shortening of G1 which decreases the cytoplasmic growth rate per cell cycle, whereas in the latter case this rate is increased due to G1 prolongation. This is consistent with our hypothesis according to which the cytoplasmic/nuclear ratio must increase to a certain level to induce end stage differentiation and cell cycle arrest. A new basis of this hypothesis is the fact that end stage differentiation requires large quantities of membranous cytoplasmic structures that the cells are unable to produce de novo. Embryonic cells, however, possess only few of these structures. The only feasible way to multiply these structures is by growing more cytoplasm per cell cycle than needed for a doubling so that successively, the level of the cytoplasmic/nuclear ratio is reached that is required for differentiation. A consequence is that the cytoplasmic growth rate per cell cycle determines the number of amplification divisions. We suggest that the differentiation signal may be triggered when a differentiation-preventing protein (for example Bcl-2) is diluted out by the expansion of cytoplasmic membrane structures, thus simultaneously determining the cell size. The intracellular timer and extracellular signals cooperate in adjusting cell production to the organism's need and in determining when and how the cells respond to extracellular signals or transmit extracellular signals.

Animals↗
PubMed2003

The cytosolic chaperonin CCT associates to cytoplasmic microtubular structures during mammalian spermiogenesis and to heterochromatin in germline and somatic cells.

Spermiogenesis, the haploid phase of spermatogenesis, is characterised by a dramatic cytodifferentiation of spermatids. The two major steps, nuclear shaping and cytoplasmic reorganisation of the organelles, rely on an extensive remodelling of the microtubule cytoskeleton. Folding of alpha- and beta-tubulin is mediated by the cytoplasmic chaperonin containing TCP-1 (CCT), highly expressed in testis. We studied CCT cellular distribution throughout spermatogenesis by immunofluorescence and immunoelectron microscopy. We unveil two main cytoplasmic localisations for CCT: at the centrosome and at the microtubules of the manchette, a structure unique to male germ cells. Both structures are essential for spermatid differentiation and may require CCT function. Although CCT is essentially cytoplasmic, a few reports suggest that a subset may have a nuclear localisation. We demonstrate that in the nucleus of germline and somatic cells, part of CCT associates to heterochromatin. In interphase cells, CCT seems generally confined to constitutive heterochromatin. Nevertheless, in condensing nucleus of future spermatozoon, it is also associated with chromatin undergoing compaction. Finally, in fully-condensed mitotic chromosomes, CCT is located all along the chromosomes. Our finding that CCT is associated with constitutive heterochromatin and to compacting chromatin raises the possibility that it may be implicated in maintenance and remodelling of heterochromatin.

Animals↗
PubMed1969

Similarities and Differences in the Development of Laboratory Strains and Freshly Isolated Strains of Herpes Simplex Virus in HEp-2 Cells: Electron Microscopy.

HEp-2 cells infected with two laboratory strains (mP and MP) and two freshly isolated strains (F and G) of herpes simplex virus were fixed at intervals between 4 and 50 hr postinfection and sectioned, and were then examined with the electron microscope. These studies revealed the following. (i) All four strains caused identical segregation of nucleoli and aggregation of host chromosomes at the nuclear membrane. (ii) The development of MP virus could not be differentiated from that of its parent mP strain. (iii) There were quantitative differences between laboratory (mP) and freshly isolated (F) type 1 strains. Thus, cells infected with F contained numerous nuclear crystals of nucleocapsids and relatively few cytoplasmic structures containing enveloped nucleocapsids. Conversely, cells infected with mP or with MP virus contained numerous cytoplasmic structures with enveloped nucleocapsids and relatively few nuclear crystals of nucleocapsids. (iv) There were qualitative differences between type 2 strain (G) isolated from genital lesions and type 1 strains. Thus, cells infected with the G strain contain numerous filaments in nuclei and unenveloped and partially enveloped nucleocapsids in the cytoplasm. Of particular interest is the finding that cytoplasmic membranes in apposition to nucleocapsids were thickened and bent as if they were enveloping the particle. The significance of the qualitative differences in the development of the four strains is discussed.

Journal Article↗