Extraction of flagellum basal body complexes from Buchnera aphidicola, an endosymbiont of aphids v1 (protocols.io.bs5wng7e)

Buchnera aphidicola is an intracellular bacterial symbiont of aphids and maintains a small genome of only 600 kbps. Buchnera is thought to maintain only genes relevant to the symbiosis with its aphid host. Curiously, the Buchnera genome contains gene clusters coding for flagellum basal body structural proteins and for flagellum type III export machinery. These structures have been shown to be highly expressed and present in large numbers on Buchnera cells. No recognizable pathogenicity factors or secreted proteins have been identified in the Buchnera genome, and the relevance of this protein complex to the symbiosis is unknown. Here, we show isolation of Buchnera flagellum basal body proteins from the cellular membrane of Buchnera, confirming the enrichment of flagellum basal body proteins relative to other proteins in the Buchnera proteome. This will facilitate studies of the structure and function of the Buchnera flagellum structure, and its role in this model symbiosis.

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Isolation of the Buchnera aphidicola flagellum basal body from the Buchnera membrane

10.1101/2021.01.07.425737 ◽

2021 ◽

Author(s):

Matthew J Schepers ◽

James N Yelland ◽

Nancy A Moran ◽

David W Taylor

Keyword(s):

Basal Body ◽

Gene Clusters ◽

Cellular Membrane ◽

Structure And Function ◽

Bacterial Symbiont ◽

Structural Proteins ◽

Buchnera Aphidicola ◽

Large Numbers ◽

And Function ◽

Aphid Host

Buchnera aphidicola is an intracellular bacterial symbiont of aphids and maintains a small genome of only 600 kbps. Buchnera is thought to maintain only genes relevant to the symbiosis with its aphid host. Curiously, the Buchnera genome contains gene clusters coding for flagellum basal body structural proteins and for flagellum type III export machinery. These structures have been shown to be highly expressed and present in large numbers on Buchnera cells. No recognizable pathogenicity factors or secreted proteins have been identified in the Buchnera genome, and the relevance of this protein complex to the symbiosis is unknown. Here, we show isolation of Buchnera flagella from the cellular membrane of Buchnera, confirming the enrichment of flagellum proteins relative to other proteins in the Buchnera proteome. This will facilitate studies of the structure and function of the Buchnera flagellum structure, and its role in this model symbiosis.

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The three-dimensional structure of frozen-hydrated left- and right-handed forms of bacterial flagellar filaments from an identical serotype

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100143110 ◽

1986 ◽

Vol 44 ◽

pp. 298-299

Author(s):

S. Trachtenberg ◽

D. J. DeRosier

Keyword(s):

Ionic Strength ◽

Basal Body ◽

Three Dimensional ◽

Dimensional Structure ◽

Protein Species ◽

Liquid Environment ◽

Bacterial Flagellum ◽

Left And Right ◽

Rotary Motor ◽

Helical Parameters

The bacterial cell is propelled through the liquid environment by means of one or more rotating flagella. The bacterial flagellum is composed of a basal body (rotary motor), hook (universal coupler), and filament (propellor). The filament is a rigid helical assembly of only one protein species — flagellin. The filament can adopt different morphologies and change, reversibly, its helical parameters (pitch and hand) as a function of mechanical stress and chemical changes (pH, ionic strength) in the environment.

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Ultrastructure of the basal body apparatus in epidermal cells of a sponge larva (Aplysilla SP: Demospongiae)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012504x ◽

1992 ◽

Vol 50 (1) ◽

pp. 928-929

Author(s):

R.L. Pinto ◽

R.M. Woollacott

Keyword(s):

Sodium Bicarbonate ◽

Epoxy Resin ◽

Basal Body ◽

Phosphate Buffer ◽

Similar Data ◽

Basal Apparatus ◽

Striated Rootlet ◽

The Common ◽

Basal Foot ◽

Sexually Mature

The basal body and its associated rootlet are the organelles responsible for anchoring the flagellum or cilium in the cytoplasm. Structurally, the common denominators of the basal apparatus are the basal body, a basal foot from which microtubules or microfilaments emanate, and a striated rootlet. A study of the basal apparatus from cells of the epidermis of a sponge larva was initiated to provide a comparison with similar data on adult sponges.Sexually mature colonies of Aplysillasp were collected from Keehi Lagoon Marina, Honolulu, Hawaii. Larvae were fixed in 2.5% glutaraldehyde and 0.14 M NaCl in 0.2 M Millonig’s phosphate buffer (pH 7.4). Specimens were postfixed in 1% OsO4 in 1.25% sodium bicarbonate (pH 7.2) and embedded in epoxy resin. The larva ofAplysilla sp was previously described (as Dendrilla cactus) based on live observations and SEM by Woollacott and Hadfield.

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Ciliary coordination in newt lung cells requires microtubule-based linkages between basal bodies: A correlative light and EM study

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100123258 ◽

1992 ◽

Vol 50 (1) ◽

pp. 570-571

Author(s):

Robert Hard ◽

Gerald Rupp ◽

Matthew L. Withiam-Leitch ◽

Lisa Cardamone

Keyword(s):

Basal Body ◽

Untreated Control ◽

Beat Frequency ◽

Primary Cultures ◽

Living Cells ◽

Power Stroke ◽

Ultrastructural Changes ◽

Lung Cells ◽

Basal Bodies ◽

Ciliated Cells

In a coordinated field of beating cilia, the direction of the power stroke is correlated with the orientation of basal body appendages, called basal feet. In newt lung ciliated cells, adjacent basal feet are interconnected by cold-stable microtubules (basal MTs). In the present study, we investigate the hypothesis that these basal MTs stabilize ciliary distribution and alignment. To accomplish this, newt lung primary cultures were treated with the microtubule disrupting agent, Colcemid. In newt lung cultures, cilia normally disperse in a characteristic fashion as the mucociliary epithelium migrates from the tissue explant. Four arbitrary, but progressive stages of dispersion were defined and used to monitor this redistribution process. Ciliaiy beat frequency, coordination, and dispersion were assessed for 91 hrs in untreated (control) and treated cultures. When compared to controls, cilia dispersed more rapidly and ciliary coordination decreased markedly in cultures treated with Colcemid (2 mM). Correlative LM/EM was used to assess whether these effects of Colcemid were coupled to ultrastructural changes. Living cells were defined as having coordinated or uncoordinated cilia and then were processed for transmission EM.

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