Shimoike, T, Taura, T, Kihara, A, Yoshihisa, T, Akiyama, Y, Cannon, K and Ito, K (1995) Product of a new gene, syd, functionally interacts with SecY when overproduced in Escherichia coli. J. Biol. Chem. 270:5519-26
A mutant form of SecY, SecY-d1, was previously suggested to sequester a component(s) of the protein translocator complex. Its synthesis from a plasmid leads to interference with protein export in Escherichia coli. SecE is a target of this sequestration, and its overproduction cancels the export interference. We now report that overexpression of another gene, termed syd, also suppresses secY-d1. The nucleotide sequence of syd predicted that it encodes a protein of 181 amino acid residues, which has been identified by overproduction, purification, and determination of the amino-terminal sequence. Cell fractionation experiments suggested that Syd is loosely associated with the cytoplasmic surface of the cytoplasmic membrane. SecY may be involved in the membrane association of Syd since the association is saturable, the extent of which depends on the overproduction of SecY. SecY is rapidly degraded in vivo unless its primary partner, SecE, is sufficiently available. Overproduction of Syd was found to stabilize oversynthesized SecY. However, Syd cannot stabilize the SecY-d1 form of SecY. Thus, in the presence of both secY+ and secY-d1, Syd increases the effective SecY+/SecY-d1 ratio in the cell and cancels the dominant interference by the latter. We also found that overproduction of Syd dramatically inhibits protein export in the secY24 mutant cell in which SecY-SecE interaction has been weakened. These results indicate that Syd, especially when it is overproduced, has abilities to interact with SecY. Possible significance of such interactions is discussed in conjunction with the apparent lack of phenotypic consequences of genetic disruption of syd.
ATP-Binding Cassette Transporters; Amino Acid Sequence; Antibodies; Bacterial Outer Membrane Proteins/isolation & purification; Bacterial Outer Membrane Proteins/metabolism; Bacterial Proteins/biosynthesis; Bacterial Proteins/isolation & purification; Bacterial Proteins/metabolism; Base Sequence; Carrier Proteins/isolation & purification; Carrier Proteins/metabolism; Chromosomes, Bacterial; Electrophoresis, Polyacrylamide Gel; Escherichia coli/metabolism; Escherichia coli Proteins; Genes, Bacterial; Genes, Suppressor; Immunoblotting; Kinetics; Maltose-Binding Proteins; Membrane Proteins/biosynthesis; Membrane Proteins/isolation & purification; Membrane Proteins/metabolism; Molecular Sequence Data; Monosaccharide Transport Proteins; Peptides/chemical synthesis; Peptides/immunology; Restriction Mapping; Sequence Homology, Amino Acid
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