Abstract
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.
Highlights
Buchnera aphidicola is an obligate endosymbiont of aphid species worldwide [1] and is a model for bacterial genome reduction, maintaining one of the smallest genomes yet discovered, only 600 kbps [2, 3]
SDS-PAGE showed sixteen bands were present after the staining procedure and their sizes corresponded to those of constituent proteins of the Buchnera flagellum basal body (S1 Fig)
We compared our samples to proteomic datasets from homogenized whole aphids, and from bacteriocytes purified from pea aphids [11]
Summary
Buchnera aphidicola is an obligate endosymbiont of aphid species worldwide [1] and is a model for bacterial genome reduction, maintaining one of the smallest genomes yet discovered, only 600 kbps [2, 3]. Though the exchange of amino acids and vitamins between the aphid host and Buchnera has been well-documented [6, 8, 9], the molecular mechanism for how these metabolites cross Buchnera membranes is unknown: Buchnera maintains a small number of genes coding for membrane transport proteins, most of which are located at the inner membrane (MdlAB, PitA, YggB, ZnuABC) [2, 10]. The permeability of the Buchnera outer membrane remains a mystery, considering the paucity of annotated transporter genes in sequenced Buchnera genomes. Genes coding for proteins localizing to the outer membrane of Buchnera include small β-barrel aquaporins (OmpA, OmpF), which allow.
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