This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lee, C Articles by Beckwith, J Search for Related Content PubMed PubMed Citation Articles by Lee, C Articles by Beckwith, J

research-article

Genetic studies on the inability of beta-galactosidase to be translocated across the Escherichia coli cytoplasmic membrane.

Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.

ABSTRACT

When a signal sequence is attached to beta-galactosidase, the normally cytoplasmic protein is unable to fully traverse the cytoplasmic membrane. We used a genetic approach to study those features of beta-galactosidase responsible for the block in translocation. By using both in vivo and in vitro techniques, fragments of beta-galactosidase were interposed between a signal sequence and alkaline phosphatase. The alkaline phosphatase acts as a sensor for any blocking effects of beta-galactosidase on export. From these studies, we show that multiple regions of beta-galactosidase contribute to its failure to be translocated. These results are most easily interpreted if the folding of beta-galactosidase or of domains of it is responsible for the block in export. In addition, in certain constructs, positively charged amino acids directly following the signal sequence interfered with export.

This article has been cited by other articles:

• Sauer, E., Merdanovic, M., Price Mortimer, A., Bringmann, G., Reidl, J. (2004). PnuC and the Utilization of the Nicotinamide Riboside Analog 3-Aminopyridine in Haemophilus influenzae. Antimicrob. Agents Chemother. 48: 4532-4541 [Abstract] [Full Text]  
• Ciocchini, A. E., Roset, M. S., Inon de Iannino, N., Ugalde, R. A. (2004). Membrane Topology Analysis of Cyclic Glucan Synthase, a Virulence Determinant of Brucella abortus. J. Bacteriol. 186: 7205-7213 [Abstract] [Full Text]  
• Cruz-Ramos, H., Cook, G. M., Wu, G., Cleeter, M. W., Poole, R. K. (2004). Membrane topology and mutational analysis of Escherichia coli CydDC, an ABC-type cysteine exporter required for cytochrome assembly. Microbiology 150: 3415-3427 [Abstract] [Full Text]  
• Neves, B. C., Mundy, R., Petrovska, L., Dougan, G., Knutton, S., Frankel, G. (2003). CesD2 of Enteropathogenic Escherichia coli Is a Second Chaperone for the Type III Secretion Translocator Protein EspD. Infect. Immun. 71: 2130-2141 [Abstract] [Full Text]  
• Zenewicz, L. A., Foulds, K. E., Jiang, J., Fan, X., Shen, H. (2002). Nonsecreted Bacterial Proteins Induce Recall CD8 T Cell Responses But Do Not Serve as Protective Antigens. J. Immunol. 169: 5805-5812 [Abstract] [Full Text]  
• Adler, J., Bibi, E. (2002). Membrane Topology of the Multidrug Transporter MdfA: Complementary Gene Fusion Studies Reveal a Nonessential C-Terminal Domain. J. Bacteriol. 184: 3313-3320 [Abstract] [Full Text]  
• Møller, P. L., Jørgensen, F., Hansen, O. C., Madsen, S. M., Stougaard, P. (2001). Intra- and Extracellular {beta}-Galactosidases from Bifidobacterium bifidum and B. infantis: Molecular Cloning, Heterologous Expression, and Comparative Characterization. Appl. Environ. Microbiol. 67: 2276-2283 [Abstract] [Full Text]  
• Feilmeier, B. J., Iseminger, G., Schroeder, D., Webber, H., Phillips, G. J. (2000). Green Fluorescent Protein Functions as a Reporter for Protein Localization in Escherichia coli. J. Bacteriol. 182: 4068-4076 [Abstract] [Full Text]  
• Piazza, F., Tortosa, P., Dubnau, D. (1999). Mutational Analysis and Membrane Topology of ComP, a Quorum-Sensing Histidine Kinase of Bacillus subtilis Controlling Competence Development. J. Bacteriol. 181: 4540-4548 [Abstract] [Full Text]  
• Fekkes, P., Driessen, A. J. M. (1999). Protein Targeting to the Bacterial Cytoplasmic Membrane. Microbiol. Mol. Biol. Rev. 63: 161-173 [Abstract] [Full Text]  
• Debarbieux, L., Beckwith, J. (1998). The reductive enzyme thioredoxin 1 acts as an oxidant when it is exported to the Escherichia coli periplasm. Proc. Natl. Acad. Sci. USA 95: 10751-10756 [Abstract] [Full Text]  
• Stephenson, K., Harwood, C. R. (1998). Influence of a Cell-Wall-Associated Protease on Production of alpha -Amylase by Bacillus subtilis. Appl. Environ. Microbiol. 64: 2875-2881 [Abstract] [Full Text]  
• Zimmann, P., Puppe, W., Altendorf, K. (1995). Membrane Topology Analysis of the Sensor Kinase KdpD of Escherichia coli. J. Biol. Chem. 270: 28282-28288 [Abstract] [Full Text]