scholarly journals BASAL BODIES OF BACTERIAL FLAGELLA IN PROTEUS MIRABILIS

1966 ◽  
Vol 31 (3) ◽  
pp. 603-618 ◽  
Author(s):  
Judith F. M. Hoeniger ◽  
Woutera van Iterson ◽  
Eva Nijman van Zanten
Keyword(s):  
Plasma Membrane ◽  
Proteus Mirabilis ◽  
Basal Bodies ◽  
Previous Suggestion ◽  
Potassium Tellurite ◽  
Bacterial Flagella ◽  
Left Behind ◽  
Potassium Phosphotungstate ◽  
Cytoplasmic Content

This paper investigates further the question of whether the flagella of Proteus mirabilis emerge from basal bodies. The bacteria were grown to the stage of swarmer differentiation, treated lightly with penicillin, and then shocked osmotically. As a result of this treatment, much of the cytoplasmic content and also part of the plasma membrane were removed from the cells. When such fragmented organisms were stained negatively with potassium phosphotungstate, the flagella were found to be anchored—often by means of a hook—in rounded structures approximately 50 mµ wide, thus confirming Part I of our study. In these rounded structures a more brilliant dot was occasionally observed, which we interpret as being part of the basal granule. A prerequisite for the demonstration of the basal granules within the cells was, however, the removal of both the cytoplasm and the plasma membrane from their vicinity. In some experiments, the chondrioids were "stained" positively by the incorporation into them of the reduced product of potassium tellurite. The chondrioids were here observed to be more or less circular areas from which rodlike structures extended. The chondrioids adhered so firmly to the plasma membrane that they were carried away with it during its displacement by osmotic shocking, while the basal bodies were left behind. This observation disproves our previous suggestion that the flagella might terminate in the chondrioids. The basal bodies often occur in pairs, which suggest that they could be self-reproducing particles.

scholarly journals BASAL BODIES OF BACTERIAL FLAGELLA IN PROTEUS MIRABILIS

1966 ◽  
Vol 31 (3) ◽  
pp. 585-602 ◽  
Author(s):  
Woutera van Iterson ◽  
Judith F. M. Hoeniger ◽  
Eva Nijman van Zanten
Keyword(s):  
Proteus Mirabilis ◽  
Cell Walls ◽  
Basal Bodies ◽  
Distilled Water ◽  
Proteus Vulgaris ◽  
Fixation Technique ◽  
Previous Suggestion ◽  
Thin Sections ◽  
Bacterial Flagella ◽  
Whole Mount

Years ago (16, 18, 19), in a study of shadowed preparations of Proteus vulgaris that had been autolyzed in the cold, the observation was made that the flagella arose from basal bodies. However, recently (3, 7, 24, 33) doubt has been cast on the conclusion that the flagella of bacteria emerge from sizable basal bodies. This problem has, therefore, been reinvestigated with actively developing cultures of Proteus mirabilis, the cell walls of which had been expanded slightly by exposure to penicillin. Two techniques were applied: ultramicrotomy, and negative staining of whole mount preparations. This paper deals with the thin sections of bacteria after the usual fixation technique had been altered slightly: the cells were embedded in agar prior to their fixation and further processing. The flagella then remained attached to the cells and were seen to extend between the cell wall and the plasma membrane. Occasionally, the flagella appeared to be anchored in the cell by means of a hook-shaped ending. In sections of cells rich in cytoplasm, the basal bodies are particularly difficult to visualize due to their small size (25 to 45 mµ) and the lack of properties that would enable one to distinguish them from the ribonucleoprotein structures; in addition, their boundary appears to be delicate. However, when the cytoplasm is sparse in the cells, either naturally or as a result of osmotic shocking in distilled water, the flagella can be observed to emerge from rounded structures approximately 25 to 45 mµ wide. Contrary to a previous suggestion (21), the flagella do not terminate in the peripheral sites of reduced tellurite, i.e. the chondrioids. The observations in this part of the study agree with those described in the following paper (15) dealing with negatively stained preparations.

Fine-Structural Aspects of Fertilization in Chlamydomonas Reinhardi

1968 ◽  
Vol 3 (1) ◽  
pp. 115-128
Author(s):  
I. FRIEDMANN ◽  
A. L. COLWIN ◽  
LAURA H. COLWIN
Keyword(s):  
Plasma Membrane ◽  
Mating Type ◽  
Starch Content ◽  
The Body ◽  
Structural Basis ◽  
Basal Bodies ◽  
Cytoplasmic Organelle ◽  
Animal Phyla ◽  
Small Structure ◽  
Specialized Structure

Gametes of C. reinhardi lack the cell wall which vegetative cells possess. Just below the cell apex gametes form a fertilization tubule which is up to 2 µ long and 0.2 µ in diameter; its plasma membrane and that of the apex have slender tubular projections. At the base of the fertilization tubule regularly lies the choanoid body, a collar-shaped cytoplasmic organelle; the plasma membrane overlying the body appears as an electron-dense ring. Gametes possess two ‘free’ basal bodies in addition to the basal bodies of the two flagella. In the initial stage of union the conjugating cells are connected by the fertilization tubule whose plasma membrane is continuous with that of both copulants. At one end of the tubule lies a conspicuous choanoid body, but at the other end is a small structure which possibly is a homologue of the choanoid body. Subsequently, the fertilization tubule shortens and widens until finally no tubule exists and the apical ends of the two protoplasts adjoin. The merging cells then bend like a jack-knife and lateral alignment of the protoplasts occurs. This four-flagellated zygote becomes motile at about the time when the flagellar bases of the former gametes seem to approach each other and when fibrillar elements of the flagellar roots come into contact. In the motile zygote the nuclei do not fuse but remain ensheathed in the cup-shaped plastids of the two gametes. A mating of plus (+) and minus (-) strains cultured, respectively, for high and low starch content suggested that gametes of only the plus (+) mating type contain the choanoid body. Since it appears that the gamete containing the choanoid body also produces the fertilization tubule, it is inferred that gametes of only the plus (+) mating type produce the fertilization tubule. Should further investigation support this inference, it would be established that there is a structural basis for designating the plus (+) mating type as male and the minus (-) type as female. Fertilization involves fusion of the gamete membranes through the mediation of a specialized structure (the fertilization tubule) and in this respect there are similarities to certain aspects of fertilization in animal phyla. The relation of the fertilization tubule to the protoplasmic bridge of other species of Chlamydomonas is discussed.

scholarly journals A CYTOCHEMICAL LOCALIZATION OF REDUCTIVE SITES IN A GRAM-POSITIVE BACTERIUM

1964 ◽  
Vol 20 (3) ◽  
pp. 361-375 ◽  
Author(s):  
Woutera van Iterson ◽  
W. Leene
Keyword(s):  
Plasma Membrane ◽  
Cell Periphery ◽  
Gram Positive ◽  
Cytochemical Localization ◽  
Potassium Tellurite ◽  
Thin Sections ◽  
Gram Positive Bacteria ◽  
Cytoplasmic Membranes ◽  
Reducing Activity ◽  
The One

In bacteria the exact location of a respiratory enzyme system comparable to that of the mitochondria of other cells has remained uncertain. On the one hand, the existence of particulate "bacterial mitochondria" has been advocated (Mudd); on the other hand, important enzymes of the respiratory chain were recovered in the cytoplasmic membranes associated with some granular material (Weibull). In order to gain insight into this question, sites of reducing activity were localized in thin sections of bacteria using the reduction of potassium tellurite as an indicator. When this salt was added to the culture medium of Bacillus subtilis, it turned out that in this Gram-positive organism the reduced product is strictly bound at two sites, and that the plasma membrane does not materially gain in electron opacity through deposition of the reduced product. The reduction product is found on or in the membranes of particular organelles, which may possibly be regarded as the mitochondrial equivalents in Gram-positive bacteria, and which are sometimes seen connected to the plasma membrane. The second location is in thin rod-like elements at the cell periphery, possibly the sites from which the flagella emerge.

scholarly journals MinD-like ATPase FlhG effects location and number of bacterial flagella during C-ring assembly

2015 ◽  
Vol 112 (10) ◽  
pp. 3092-3097 ◽  
Author(s):  
Jan S. Schuhmacher ◽  
Florian Rossmann ◽  
Felix Dempwolff ◽  
Carina Knauer ◽  
Florian Altegoer ◽  
...  
Keyword(s):  
Cell Division ◽  
Structural Diversity ◽  
Basal Bodies ◽  
Independent Manner ◽  
Bacterial Flagella ◽  
Division Site ◽  
Ring Assembly ◽  
Species Specific ◽  
Regular Patterns

The number and location of flagella, bacterial organelles of locomotion, are species specific and appear in regular patterns that represent one of the earliest taxonomic criteria in microbiology. However, the mechanisms that reproducibly establish these patterns during each round of cell division are poorly understood. FlhG (previously YlxH) is a major determinant for a variety of flagellation patterns. Here, we show that FlhG is a structural homolog of the ATPase MinD, which serves in cell-division site determination. Like MinD, FlhG forms homodimers that are dependent on ATP and lipids. It interacts with a complex of the flagellar C-ring proteins FliM and FliY (also FliN) in the Gram-positive, peritrichous-flagellated Bacillus subtilis and the Gram-negative, polar-flagellated Shewanella putrefaciens. FlhG interacts with FliM/FliY in a nucleotide-independent manner and activates FliM/FliY to assemble with the C-ring protein FliG in vitro. FlhG-driven assembly of the FliM/FliY/FliG complex is strongly enhanced by ATP and lipids. The protein shows a highly dynamic subcellular distribution between cytoplasm and flagellar basal bodies, suggesting that FlhG effects flagellar location and number during assembly of the C-ring. We describe the molecular evolution of a MinD-like ATPase into a flagellation pattern effector and suggest that the underappreciated structural diversity of the C-ring proteins might contribute to the formation of different flagellation patterns.

SITES OF "GLYCINE OXIDASE" ACTIVITY IN PROTEUS VULGARIS

1967 ◽  
Vol 13 (5) ◽  
pp. 551-556
Author(s):  
M. V. Nermut
Keyword(s):  
Plasma Membrane ◽  
Oxidase Activity ◽  
Proteus Vulgaris ◽  
Potassium Tellurite ◽  
Close Vicinity ◽  
Ultrathin Sections ◽  
Glycine Oxidase

The location of glycine oxidase activity in Proteus vulgaris was investigated using potassium tellurite and the technique of ultrathin sections. In 95% of cases, the tellurium deposits were found in the close vicinity of the plasma membrane, presumably at the inner side of it.

scholarly journals A "MICROTUBULE" IN A BACTERIUM

1967 ◽  
Vol 32 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Woutera van Iterson ◽  
Judith F. M. Hoeniger ◽  
Eva Nijman van Zanten
Keyword(s):  
Proteus Mirabilis ◽  
Normal Cell ◽  
Osmotic Shock ◽  
Hollow Core ◽  
Cylindrical Wall ◽  
Thin Sections ◽  
Whole Mount ◽  
Potassium Phosphotungstate ◽  
First Time ◽  
Cell Constituent

A study of the anchorage of the flagella in swarmers of Proteus mirabilis led to the incidental observation of microtubules. These microtubules were found in thin sections and in whole mount preparations of cells from which most of the content had been released by osmotic shock before staining negatively with potassium phosphotungstate (PTA). The microtubules are in negatively stained preparations about 200 A wide, i.e. somewhat thicker than the flagella (approximately 130 A). They are thus somewhat thinner than most microtubules recorded for other cells. They are referred to as microtubules because of their smooth cylindrical wall, or cortex, surrounding a hollow core which is readily filled with PTA when stained negatively. Since this is probably the first time that such a structure is described inside a bacterium, we do not know for certain whether it represents a normal cell constituent or an abnormality, for instance of the type of "polysheaths" (16).

scholarly journals THE FINE STRUCTURE AND FUNCTION OF THE CONTRACTILE AXOSTYLES OF CERTAIN FLAGELLATES

1965 ◽  
Vol 24 (3) ◽  
pp. 387-400 ◽  
Author(s):  
A. V. Grimstone ◽  
L. R. Cleveland
Keyword(s):  
Plasma Membrane ◽  
Fine Structure ◽  
Structure And Function ◽  
Basal Bodies ◽  
Structural Components ◽  
Cross Links ◽  
Cilia And Flagella ◽  
Fine Structure And Function ◽  
And Function ◽  
Regular Arrays

The axostyles of the flagellates Oxymonas, Saccinobaculus, and Notila are large ribbon-shaped structures which undulate actively in the cytoplasm. The form of their movements is described and illustrated. Axostyles consist of regular arrays of longitudinal fibres, the number of which varies between 100 and 5000 in different species. The fibres are about 240 A in diameter, apparently hollow, regularly cross-banded with a periodicity of about 150 A, and connected by delicate cross-links, also at regular intervals of about 150 A. They resemble very closely the central fibres of cilia and flagella. No other structural components are present, except at the anterior end, where the fibres are attached to one or more basal bodies, and at the posterior tip, where they are anchored to the plasma membrane. The relevance of the findings to an understanding of the mechanism of ciliary and flagellar movements is discussed.

Galectin-3-mediated adherence ofProteus mirabilisto Madin-Darby canine kidney cells

2001 ◽  
Vol 79 (6) ◽  
pp. 783-788 ◽  
Author(s):  
Eleonora Altman ◽  
Blair A Harrison ◽  
Roger K Latta ◽  
Kok K Lee ◽  
John F Kelly ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Plasma Membrane ◽  
Proteus Mirabilis ◽  
Mdck Cells ◽  
Bacterial Adherence ◽  
Madin Darby Canine Kidney ◽  
Galectin 3 ◽  
Tract Infections ◽  
Darby Canine Kidney ◽  
Canine Kidney

Proteus mirabilis is an important cause of urinary tract infections (UTIs) and can result in acute pyelonephritis. Proteus mirabilis expresses several, morphologically distinct, fimbrial species, and previous studies have shown that the nonagglutinating fimbriae (NAF) mediate bacterial adherence to a number of cell lines, including Madin-Darby canine kidney (MDCK) cells. Immunoblot overlay analysis of the plasma membrane fraction from MDCK cells with purified NAF revealed a 34-kDa band, which has been analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Database search identified galectin-3 as a potential protein candidate. Immunocytochemical assay of MDCK cells with a galectin-3-specific monoclonal antibody, anti-Mac-2, confirmed its presence on the plasma membrane extracellular surface. Preincubation of P. mirabilis with anti-Mac-2 monoclonal antibodies, specific for galectin-3, resulted in the inhibition of bacterial binding to MDCK cells. These data suggest a role for galectin-3, interacting with appropriately glycosylated surface receptors and P. mirabilis fimbriae, as a mediator of bacterial adherence in vitro.Key words: bacterial adherence, fimbriae, galectin-3, mass spectrometry.

scholarly journals The ternary complex CEP90, FOPNL and OFD1 specifies the future location of centriolar distal appendages, and promotes their assembly

2021 ◽  
Author(s):  
Pierrick Le Borgne ◽  
Marine Hélène Laporte ◽  
Logan Greibill ◽  
Michel Lemullois ◽  
Mebarek Temagoult ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Plasma Membrane ◽  
Transition Zone ◽  
Basal Body ◽  
Mammalian Cells ◽  
Ternary Complex ◽  
Basal Bodies ◽  
Sequence Of Events ◽  
The Future ◽  
High Conservation

Cilia assembly starts with centriole to basal body maturation, migration to the cell surface and docking to the plasma membrane. The basal body docking process involves the interaction of both the distal end of the basal body and the transition fibers (or mature distal appendages), with the plasma membrane. During this process, the transition zone assembles and forms the structural junction between the basal body and the nascent cilium. Mutations in numerous genes involved in basal body docking and transition zone assembly are associated with the most severe ciliopathies, highlighting the importance of these events in cilium biogenesis. The conservation of this sequence of events across phyla is paralleled by a high conservation of the proteins involved. We identified CEP90 by BioID using FOPNL as a bait. Ultrastructure expansion microscopy showed that CEP90, FOPNL and OFD1 localized at the distal end of both centrioles/basal bodies in Paramecium and mammalian cells. These proteins are recruited early after duplication on the procentriole. Finally, functional analysis performed both in Paramecium and mammalian cells demonstrate the requirement of this complex for distal appendage assembly and basal body docking. Altogether, we propose that this ternary complex is required to determine the future position of distal appendages

scholarly journals Initial ciliary assembly in Chlamydomonas requires Arp2/3-dependent recruitment from a ciliary protein reservoir in the plasma membrane

2020 ◽  
Author(s):  
Brae M Bigge ◽  
Nicholas E Rosenthal ◽  
David Sept ◽  
Courtney M Schroeder ◽  
Prachee Avasthi
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Long Range ◽  
Protein Targeting ◽  
Basal Bodies ◽  
Cell Periphery ◽  
Filamentous Actin ◽  
Actin Networks ◽  
Ciliary Protein ◽  
Broad Approach

ABSTRACTCilia are organelles important for signaling and motility. They are composed of microtubules ensheathed in plasma membrane. The mechanisms related to ciliogenesis also require another cytoskeletal element, actin, which has been shown to be important for organizing the basal bodies and transition zone at the base of cilia and for short- and long-range trafficking. However, most studies of actin’s role in ciliogenesis have taken a broad approach by knocking out all filamentous actin until now. Here, we more delicately dissect the interplay between actin and cilia by specifically focusing on actin networks nucleated by the Arp2/3 complex in Chlamydomonas. We find that knocking out Arp2/3-mediated actin networks dramatically impairs ciliary assembly and maintenance in these cells, and these defects are due to a problem with incorporation and gating of existing ciliary proteins, particularly in the early stages of assembly. We also show that cells lacking the Arp2/3 complex have more dramatic defects in ciliary maintenance using material from non-Golgi sources. Finally, we find relocalization of a ciliary membrane protein from the cell periphery to the cilia by internalization is dependent on actin and the Arp2/3 complex. Based on these results, we propose a new model of ciliary protein targeting during early ciliogenesis in which proteins previously targeted from the Golgi to the plasma membrane are reclaimed from this reservoir by Arp2/3-mediated networks.

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