Ultrastructure of the attachment of Pyrsonympha to the hind-gut wall of Reticulitermes tibialis

Protozoa of the genus Pyrsonympha are found attached to the cuticular lining of the hind-gut wall of the termite Reticulitermes tibialis by a specialized anterior structure referred to here as the ‘attachment organelle’. The structure of the hind-gut, the attachment organelle, and the nature of the attachment are described using light and electron microscopy. The attachment organelle is a highly folded membranous structure containing large 20 to 26 nm filaments. Although no unambiguous functional significance can presently be assigned to this attachment, it may serve to prevent loss of the protozoa from the intestinal tract or it may be involved in the nutritional symbiosis between termite and protozoa.     

Structural changes in proteins of the bacteriophage T4 basal plate resulting from the attachment of long fibrils

The effect of the attachment of long tail fibers on the structure of proteins of the bacteriophage T4 baseplate was studied by digital processing of electron microscopic images. The attachment of the long fibers was found to result in dramatical changes of the proteins of the baseplate plag, while the wedges, to which the long fibers are attached, undergo only slight changes. We studied the baseplates with one to six attached fibers and found that the attachment of one fiber resulted in the change of the entire baseplate, although the wedge located in the vicinity of the fiber attachment changed to a greater extent. Only after the attachment of three and more fibers the changes of the same kind occurred through the entire baseplate.     

Analysis of the attachment of replicating DNA to a nuclear matrix in mammalian interphase nuclei.

The attachment of replicating DNA to a rapidly sedimenting nuclear structure was investigated by digestion with various nucleases. When DNA was gradually removed by DNase I, pulse label incorporated during either 1 min or during 1 hour in the presence of arabinosylcytosine, remained preferentially attached to the nuclear structure. Single strand specific digestion by nuclease S1 or staphylococcal nuclease at low concentrations caused a release of about 30% of the pulse label, without significantly affecting the attachment of randomly labelled DNA. The released material had a low sedimentation coefficient and contained most of the Okasaki fragments. The remaining pulse label was less accessible to further digestion by double strand specific nuclease activity than the bulk DNA. The results suggest that an attachment of the replication fork to the nuclear structure occurs at sites behind but close to the branch point.     

Ultrastructure of the membrane attachment sites of the extrusomes of Ciliophrys marina and Heterophrys marina (Actinopoda)

Summary The actinopods Ciliophrys marina and Heterophrys marina both have membrane bounded extrusomes attached to their cellular and axopodial membranes. The extrusomes of C. marina, the muciferous bodies, are fairly simple in structure and contain a homogeneous osmiophilic substance. Their attachment site is characterized by a rectangular array of freeze fracture particles in the cell membrane. The extrusomes of H. marina, the conicysts, are more complex and contain a two-part osmiophilic body. The attachment site of conicysts is characterized by a rosette of 8 freeze fracture particles very similar to the 9-particle rosette found at the mucocyst attachment sites in Tetrahymena. Furthermore, intracytoplasmic bridges connect the conicyst and cell membrane faces, and a specialized fibrillar structure is found on the cell membrane in the region of conicyst attachment. The various possible roles for such particle arrays are discussed and their presence in virtually all extrusomes is predicted.Supported by USPHS GM01021 and HL13849     

Ultrastructure of the scolex of Orygmatobothrium schmittii (Cestoda: Phyllobothriidea)

The ultrastructure of the scolex of Orygmatobothrium schmittii (Cestoda: Phyllobothriidae) was studied using histochemistry, scanning, and transmission electron microscopy. The central bothridial structure resulted in a glandulomuscular organ formed by a mass of syncytial glands and radial muscles, with glycoprotein secretions potentially adhesive. Among the sensory receptors found on the scolex, a particular type was found surrounding the glandulomuscular organ, which might be related in the regulation of the secretions. The internal structure of the microtriches revealed a diversity of configurations according to their morphotype and distribution on the scolex. Microtriches with larger caps are thought to be useful for attachment purposes. In addition, the thick bounding membranes of the attachment organs and the circular musculature in the bothridia, seem to aid to the attachment of the scolex to the mucosa of the host.     

Observations on the structure of larval attachment organs in three species of gymnotiforms (Teleostei: Ostariophysi)

Attachment organs are known in certain teleosts but have not been reported in gymnotiforms. This study demonstrates the presence and describes the structure of attachment organs in three species of gymnotiforms: the apteronotids Apteronotus albifrons and A. leptorhynchus , and the rhamphichthyid Rhamphichthys sp. Attachment organs are present only during the first few days after hatching. Organs were studied with light microscopy, SEM, and TEM. In all three species attachment organs are restricted to the head region. They consist of numerous individual attachment cells and modified epithelial cells distributed on the frontal, lateral, and dorsal head area. Only the apical parts of the attachment cells reach the body surface between the epithelium of the epidermis. Attachment cells contain considerable amounts of rough ER and Golgi vesicles filled a the glutinous substance. Epithelial cells in the area in which attachment cells are developed differ in structure from those on the rest of the body and form a functional complex with the attachment cells. In the two Apteronotus species they possess 4–10 μm long projections that are located in the cell centre in A. albifrons , but are slightly shifted to one side of the cells in A. leptorhynchus . Epithelial cells in the organ area of Rhamphichthys sp. lack any projections and resemble cells of the trunk region. However, their pattern of microridges on the outer cell surface is more pronounced and conspicuous. Projections along with modified microridges may aid in distributing and focusing adhesive substances released by attachment cells, and thus may enhance adhesive properties.     

The fine structure of the interrelationship of cells in the human epidermis

In the present investigation an analysis has been made of the fine structure of the interrelationships of cells in human forearm epidermis by means of the electron microscope. The "intercellular bridges," here called attachment zones, are more complex than has previously been recognized. It is shown that dense oval thickenings, called attachment plaques, appear in apposed areas of adjacent epidermal cell membranes. The tonofibrils terminate at the internal face of the attachment plaque and do not traverse the 300 A distance between apposed plaques. Seven intervening layers of unidentified substance occupy the space between attachment plaques. The attachment zones appear in all of the classical histological layers of the epidermis. The portions of epidermal cell membrane not involved in intercellular attachments have extensive surface area resulting from plication of the membrane, and its further modification to form microvilli. The possible functional significance of these observations is discussed. Prior observations concerning the basement membrane of epidermis are confirmed. Identification of epidermal melanocytes is achieved, the finer morphology of their dendritic processes is described, and their relationship to epidermal cells is discussed.     

A Mechanical Model for the Adhesion of Spiders to Nominally Flat Surfaces

In dry attachment systems of spiders and geckos, van der Waals forces mediate attraction between substrate and animaltarsus. In particular, the scopula of Evarcha arcuata spiders allows for reversible attachment and easy detachment to a broadrange of surfaces. Hence, reproducing the scopula’s roughness compatibility while maintaining anti-bunching features and dirtparticle repellence behavior is a central task for a biomimetic transfer to an engineered model. In the present work we model thescopula of E. arcuata from a mechano-elastic point of view analyzing the influence of its hierarchical structure on the attachmentbehavior. By considering biological data of the gecko and spider, and the simulation results, the adhesive capabilities of thetwo animals are compared and important confirmations and new directives in order to reproduce the overall structure are found.Moreover, a possible suggestion of how the spider detaches in an easy and fast manner is proposed and supported by the results.     

Bacterial cell attachment, the beginning of a biofilm

The ability of bacteria to attach to surfaces and develop into a biofilm has been of considerable interest to many groups in numerous industries, including the medical and food industry. However, little is understood in the critical initial step seen in all biofilm development, the initial bacterial cell attachment to a surface. This initial attachment is critical for the formation of a bacterial biofilm, as all other cells within a biofilm structure rely on the interaction between surface and bacterial cell for their survival. This review examines what are believed to be some of the most important aspects involved in bacterial attachment to a surface.     

Characterization of DNA pattern in the site of permanent attachment to the nuclear matrix located in the vicinity of replication origin

The permanent sites of DNA attachment to the nuclear matrix in the domain of chicken alpha-globin genes originally found in erythrocyte nuclei have also been shown to exist in sperm and cultured fibroblast cells. A primary structure of a 1.7 kb fragment located in 5'-upstream region of chicken alpha-globin gene domain and containing both replication origin and permanent nuclear matrix attachment site has been determined. It was found to possess homologies with papovaviral replication origins and contain short internal repeats and GC-rich motifs.     

Biological activities of homologous loop regions in the laminin alpha chain G domains

Laminin alpha chains (alpha1-alpha5 chains) have diverse chain-specific biological functions. The LG4 modules of laminin alpha chains consist of a 14-stranded beta-sheet (A-N) sandwich structure. Several biologically active sequences have been identified in the connecting loop regions. Here, we evaluated the biological activities of the loop regions of the E and F strands in the LG4 modules using five homologous peptides from each of the mouse alpha chains (EF-1: DYATLQLQEGRLHFMFDLG, alpha1 chain 2747-2765; EF-2: DFGTVQLRNGFPFFSYDLG, alpha2 chain 2808-2826; EF-3: RDSFVALYLSEGHVIFALG, alpha3 chain 2266-2284; EF-4: DFMTLFLAHGRLVFMFNVG, alpha4 chain 1511-1529; EF-5: SPSLVLFLNHGHFVAQTEGP, alpha5 chain 3304-3323). These homologous peptides showed chain-specific cell attachment and neurite outgrowth activities. Well organized actin stress fibers and focal contacts with vinculin accumulation were observed in fibroblasts attached on EF-1, whereas fibroblasts on EF-2 and EF-4 showed filopodia with ruffling. Fibroblast attachment to EF-2 and EF-4 was mediated by syndecan-2. In contrast, EF-1 promoted alpha2beta1 integrin-mediated fibroblast attachment and inhibited fibroblast attachment to a recombinant laminin alpha1 chain LG4-5. The receptors for EF-3 and EF-5 are unknown. Further, when the active core sequence of EF-1 was cyclized, utilizing two additional cysteine residues at both the N and C termini through a disulfide bridge, the cyclic peptide significantly enhanced integrin-mediated cell attachment. These results indicate that integrin-mediated cell attachment to the EF-1 sequence is conformation-dependent and that the loop structure is important for the activity. The homologous peptides, which promote either integrin- or syndecan-mediated cell attachment, may be useful for understanding the cell type- and chain-specific biological activities of the laminins.     

Novel structural components of the ventral disc and lateral crest in Giardia intestinalis

Giardia intestinalis is a ubiquitous parasitic protist that is the causative agent of giardiasis, one of the most common protozoan diarrheal diseases in the world. Giardia trophozoites attach to the intestinal epithelium using a specialized and elaborate microtubule structure, the ventral disc. Surrounding the ventral disc is a less characterized putatively contractile structure, the lateral crest, which forms a continuous perimeter seal with the substrate. A better understanding of ventral disc and lateral crest structure, conformational dynamics, and biogenesis is critical for understanding the mechanism of giardial attachment to the host. To determine the components comprising the ventral disc and lateral crest, we used shotgun proteomics to identify proteins in a preparation of isolated ventral discs. Candidate disc-associated proteins, or DAPs, were GFP-tagged using a ligation-independent high-throughput cloning method. Based on disc localization, we identified eighteen novel DAPs, which more than doubles the number of known disc-associated proteins. Ten of the novel DAPs are associated with the lateral crest or outer edge of the disc, and are the first confirmed components of this structure. Using Fluorescence Recovery After Photobleaching (FRAP) with representative novel DAP::GFP strains we found that the newly identified DAPs tested did not recover after photobleaching and are therefore structural components of the ventral disc or lateral crest. Functional analyses of the novel DAPs will be central toward understanding the mechanism of ventral disc-mediated attachment and the mechanism of disc biogenesis during cell division. Since attachment of Giardia to the intestine via the ventral disc is essential for pathogenesis, it is possible that some proteins comprising the disc could be potential drug targets if their loss or disruption interfered with disc biogenesis or function, preventing attachment.     

Cyclic peptides from the loop region of the laminin alpha 4 chain LG4 module show enhanced biological activity over linear peptides

Laminins, heterotrimeric glycoproteins in the basement membrane, are involved in diverse biological activities. So far, five alpha, three beta, and three gamma chains have been identified, and at least 15 laminin isoforms exist composed of various combinations of the different three chains. The major cell-surface receptors for laminins are integrins and proteoglycans, such as dystroglycans and syndecans. Previously, we reported that synthetic peptide A4G82 (TLFLAHGRLVFM, mouse laminin alpha4 chain residues 1514-1525) showed strong cell attachment and syndecan binding activities. On the basis of the crystal structure of the LG module and sequence alignment, A4G82 is located in the connecting loop region between beta-strands E and F in the laminin alpha4 chain LG4 module. Here, we have focused on the structural importance of this E-F loop region for the biological activity of the alpha4 chain LG4 module. To determine the importance of the loop structure, we synthesized peptide A4G82X (cyclo-A4G82X, Cys-TLFLAHGRLVFX-Cys, X= norleucine), which was cyclized via disulfide bridges at both the N- and C-termini. The cyclic peptides derived from A4G82X inhibited the heparin binding activity of the alpha4 chain G domain and promoted HT-1080 cell attachment better than the corresponding linear peptides. We determined FLAHGRLVFX as a minimal sequence of cyclo-A4G82X important for cell adhesion and heparin binding using a series of truncated peptides. Moreover, HT-1080 cell attachment to the cyclic peptides was more efficiently blocked by heparin than cell attachment to the linear peptides. Furthermore, the cyclic peptides showed significantly enhanced syndecan-2-mediated cell attachment activity. These results indicate that the activity of A4G82 is highly conformation-dependent, suggesting that the E-F loop structure is crucial for its biological activity.     

Location of attachment moiety on Mycoplasma pneumoniae

Mycoplasma pneumoniae initiates infection in the human host by attachment to respiratory epithelium. The organism attaches by a specialized terminal structure. Monoclonal antibodies to an organism surface protein (P1) inhibited attachment to respiratory epithelium and were localized to the tip structure by a ferritin antibody label. The P1 protein was degraded by trypsin treatment to smaller polypeptides that possessed the same antigenic determinants as the larger P1 protein when reacted with the specific monoclonal antibody, and evidence has been provided for the existence of multiple antigenic determinants on the attachment protein.     

Making an effective switch at the kinetochore by phosphorylation and dephosphorylation

The kinetochore, the proteinaceous structure on the mitotic centromere, functions as a mechanical latch that hooks onto microtubules to support directional movement of chromosomes. The structure also brings in a number of signaling molecules, such as kinases and phosphatases, which regulate microtubule dynamics and cell cycle progression. Erroneous microtubule attachment is destabilized by Aurora B-mediated phosphorylation of multiple microtubule-binding protein complexes at the kinetochore, such as the KMN network proteins and the Ska/Dam1 complex, while Plk-dependent phosphorylation of BubR1 stabilizes kinetochore–microtubule attachment by recruiting PP2A-B56. Spindle assembly checkpoint (SAC) signaling, which is activated by unattached kinetochores and inhibits the metaphase-to-anaphase transition, depends on kinetochore recruitment of the kinase Bub1 through Mps1-mediated phosphorylation of the kinetochore protein KNL1 (also known as Blinkin in mammals, Spc105 in budding yeast, and Spc7 in fission yeast). Recruitment of protein phosphatase 1 to KNL1 is necessary to silence the SAC upon bioriented microtubule attachment. One of the key unsolved questions in the mitosis field is how a mechanical change at the kinetochore upon microtubule attachment is converted to these and other chemical signals that control microtubule attachment and the SAC. Rapid progress in the field is revealing the existence of an intricate signaling network created right on the kinetochore. Here we review the current understanding of phosphorylation-mediated regulation of kinetochore functions and discuss how this signaling network generates an accurate switch that turns on and off the signaling output in response to kinetochore–microtubule attachment.     

Conserved Fiber-Penton Base Interaction Revealed by Nearly Atomic Resolution Cryo-Electron Microscopy of the Structure of Adenovirus Provides Insight into Receptor Interaction

Adenovirus (Ad) cell attachment is initiated by the attachment of the fiber protein to a primary receptor (usually CAR or CD46). This event is followed by the engagement of the penton base protein with a secondary receptor (integrin) via its loop region, which contains an Arg-Gly-Asp (RGD) motif, to trigger virus internalization. To understand the well-orchestrated adenovirus cell attachment process that involves the fiber and the penton base, we reconstructed the structure of an Ad5F35 capsid, comprising an adenovirus type 5 (Ad5) capsid pseudotyped with an Ad35 fiber, at a resolution of approximately 4.2 Å. The fiber-penton base interaction in the cryo-electron microscopic (cryo-EM) structure of Ad5F35 is similar to that in the cryo-EM structure of Ad5, indicating that the fiber-penton base interaction of adenovirus is conserved. Our structure also confirms that the C-terminal segment of the fiber tail domain constitutes the bottom trunk of the fiber shaft. Based on the conserved fiber-penton base interaction, we have proposed a model for the interaction of Ad5F35 with its primary and secondary receptors. This model could provide insight for designing adenovirus gene delivery vectors.     

Effect of rifampin on the structure and membrane attachment of the nucleoid of Escherichia coli

Deoxyribonucleic acid (DNA) of Escherichia coli was found to be attached to the cell membrane at about 20 points. This was determined by fractionation of X-irradiated cells with the M band (magnesium-Sarkosyl crystals) technique. The number of attachment points was computed from the relationship between the amount of DNA in M bands and the number of double-strand breaks introduced by the X-ray treatment. The number of attachment points was decreased fourfold by treatment of cells with rifampin. This effect was apparently due to the action of the drug on ribonucleic acid (RNA) polymerase since the drug did not affect a mutant whose RNA polymerase is resistant to rifampin. This suggests that there may be two classes of attachment points of DNA on the membrane, some of which are removed by rifampin treatment and some which are not. Rifampin treatment also resulted in the uncondensing of isolated nucleoids and in an axial appearance of the nucleoids in ultrathin sections. The results suggest that RNA polymerase plays a role, direct or indirect, in maintaining the structure of the bacterial nucleoid and in some of its attachment to the membrane.     

Hydrodynamic effect on cell attachment to microcarriers at initial stage of microcarrier culture

An effect of intermittent agitation on cell attachment is studied. The result of cell attachment to a microcarrier was best in the case of continuous agitation. All cells attached to microcarriers under the condition of continuous agitation for 60?min. The rate of attachment was also the highest. The positive effect on cell attachment by agitation is due to the wake around the microcarrier and some changes of structure of cell membrane.