PMID:8631709
| Citation |
Dai, K, Xu, Y and Lutkenhaus, J (1996) Topological characterization of the essential Escherichia coli cell division protein FtsN. J. Bacteriol. 178:1328-34 |
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| Abstract |
Genetic and biochemical approaches were used to analyze a topological model for FtsN, a 36-kDa protein with a putative transmembrane segment near the N terminus, and to ascertain the requirements of the putative cytoplasmic and membrane-spanning domains for the function of this protein. Analysis of FtsN-PhoA fusions revealed that the putative transmembrane segment of FtsN could act as a translocation signal. Protease accessibility studies of FtsN in spheroblasts and inverted membrane vesicles confirmed that FtsN had a simple bitopic topology with a short cytoplasmic amino terminus, a single membrane-spanning domain, and a large periplasmic carboxy terminus. To ascertain the functional requirements of the N-terminal segments of FtsN, various constructs were made. Deletion of the N-terminal cytoplasmic and membrane-spanning domains led to intracellular localization of the carboxy domain, instability,and loss of function. Replacement of the N-terminal cytoplasmic and membrane-spanning domains with a membrane-spanning domain from MalG restored subcellular localization and function. These N-terminal domains of FtsN could also be replaced by the cleavable MalE signal sequence with restoration of subcellular localization and function. It is concluded that the N-terminal, cytoplasmic, and transmembrane domains of FtsN are not required for function of the carboxy domain other than to transport it to the periplasm. FtsQ and FtsI were also analyzed. |
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| Keywords |
ATP-Binding Cassette Transporters; Alkaline Phosphatase/genetics; Amino Acid Sequence; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Bacterial Proteins/ultrastructure; Biological Transport; Carrier Proteins/genetics; Carrier Proteins/metabolism; Cell Division; Endopeptidase K; Escherichia coli/growth & development; Escherichia coli Proteins; Maltose-Binding Proteins; Membrane Proteins/genetics; Membrane Proteins/metabolism; Membrane Proteins/ultrastructure; Models, Molecular; Molecular Sequence Data; Monosaccharide Transport Proteins; Peptide Mapping; Periplasmic Binding Proteins; Protein Conformation; Protein Sorting Signals; Recombinant Fusion Proteins/metabolism; Recombinant Fusion Proteins/ultrastructure; Sequence Homology, Amino Acid; Serine Endopeptidases/metabolism; Spheroplasts |
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