Assembling filamentous phage occlude pIV channels.
Filamentous phage f1 is exported from its Escherichia coli host without killing the bacterial cell. Phage-encoded protein pIV, which is required for phage assembly and secretion, forms large highly conductive channels in the outer membrane of E. coli. It has been proposed that the phage are extruded across the bacterial outer membrane through pIV channels. To test this prediction, we developed an in vivo assay by using a mutant pIV that functions in phage export but whose channel opens in the absence of phage extrusion. In E. coli lacking its native maltooligosacharride transporter LamB, this pIV variant allowed oligosaccharide transport across the outer membrane. This entry of oligosaccharide was decreased by phage production and still further decreased by production of phage that cannot be released from the cell surface. Thus, exiting phage block the pIV-dependent entry of oligosaccharide, suggesting that phage occupy the lumen of pIV channels. This study provides the first evidence, to our knowledge, for viral exit through a large aqueous channel.
Marciano
Denise K
DK
The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Russel
M
M
Simon
S M
SM
D003090:Coliphages / Q000502:physiology*; D004926:Escherichia coli / Q000648:ultrastructure / Q000821:virology; D006868:Hydrolysis; D008854:Microscopy, Electron; D021381:Protein Transport; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism
D017361:Viral Nonstructural Proteins; C119520:pIV protein, Enterobacteria phage f1