title: Both linked and unlinked mutations can alter the intracellular site of synthesis of exported proteins of Escherichia coli.
abstract: It previously has been demonstrated that synthesis of the periplasmic maltose-binding protein (MBP) and alkaline phosphatase (AP) of Eschericha coli predominantly occurs on membrane-bound polysomes. In this study, signal sequence alterations that adversely affect export of MBP and AP, resulting in their cytoplasmic accumulation as unprocessed precursors, were investigated to determine whether they have an effect on the intracellular site of synthesis of these proteins. Our findings indicate that export-defective MBP and AP are not synthesized or are synthesized in greatly reduced levels on membrane-bound polysomes. In some instances, a concomitant increase in the amount of these proteins synthesized on free polysomes was clearly discerned. We also determined the site of synthesis of MBP and AP in strains harboring mutations thought to alter the cellular secretion machinery. It was found that the presence of a prlA suppressor allele partially restored synthesis of export-defective MBP on membrane-bound polysomes. On the other hand, the absence of a functional SecA protein resulted in the synthesis of wild-type MBP and AP predominantly on free polysomes.
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000469:Alkaline Phosphatase / Q000096:biosynthesis*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis; D001692:Biological Transport; D002352:Carrier Proteins / Q000096:biosynthesis*; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D008040:Genetic Linkage; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D009154:Mutation; D010455:Peptides / Q000235:genetics; D033902:Periplasmic Binding Proteins; D011132:Polyribosomes / Q000378:metabolism; D011498:Protein Precursors / Q000096:biosynthesis; D021382:Protein Sorting Signals; D013489:Suppression, Genetic
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D010455:Peptides; D033902:Periplasmic Binding Proteins; D011498:Protein Precursors; D021382:Protein Sorting Signals; C461599:maltose transport system, E coli; D000469:Alkaline Phosphatase