J. Bacteriol., 02 1996, 1154-1161, Vol 178, No. 4
Copyright © 1996, American Society for Microbiology

Multicopy suppressors of prc mutant Escherichia coli include two HtrA (DegP) protease homologs (HhoAB), DksA, and a truncated R1pA

S Bass, Q Gu and A Christen
Department of Molecular Biology, Genentech Inc, South San Francisco, California 94080-4990, USA.

We have isolated three multicopy suppressors of the conditional lethal phenotype of a prc (tsp) null strain of Escherichia coli. One of these suppressors included two novel putative protease genes in tandem that map to 3400 kb or 72.5 centisomes on the chromosome. We propose the names hhoA and hhoB, for htrA homolog, to denote that these genes encode proteins that are 58 and 35% identical, respectively, to the HtrA (DegP) serine protease and 36% identical to each other. The HhoA and HhoB proteins are predicted to be 455 and 355 amino acids, respectively, in length. The mature HhoA protein is periplasmic in location, and amino-terminal sequencing shows that it arises following cleavage of a 27-amino-acid signal peptide. Searches of the protein and DNA databases reveal a rapidly growing family of homologous genes in a variety of other bacteria, including several which are required for virulence in their host. Deletion of the hhoAB genes shows that they are not required for viability at high temperatures like the homologous htrA but grow more slowly than wild-type strains. A second multicopy prc suppressor is the dksA (dnaK suppressor) gene, which is also a multicopy suppressor of defects in the heat shock genes dnaK, dnaJ, and grpE. The dksA gene was independently isolated as a multicopy suppressor of a mukB mutation, which is required for chromosomal partitioning. A third dosage-dependent prc suppressor includes a truncated rare lipoprotein A (rlpA) gene.

This article has been cited by other articles:

• Campbell, T. L., Ederer, C. S., Allali-Hassani, A., Brown, E. D. (2007). Isolation of the rstA Gene as a Multicopy Suppressor of YjeE, an Essential ATPase of Unknown Function in Escherichia coli. J. Bacteriol. 189: 3318-3321 [Abstract] [Full Text]  
• Zahnd, C., Pecorari, F., Straumann, N., Wyler, E., Pluckthun, A. (2006). Selection and Characterization of Her2 Binding-designed Ankyrin Repeat Proteins. J. Biol. Chem. 281: 35167-35175 [Abstract] [Full Text]  
• Kubetzko, S., Balic, E., Waibel, R., Zangemeister-Wittke, U., Pluckthun, A. (2006). PEGylation and Multimerization of the Anti-p185HER-2 Single Chain Fv Fragment 4D5: EFFECTS ON TUMOR TARGETING. J. Biol. Chem. 281: 35186-35201 [Abstract] [Full Text]  
• Yao, Y., Xie, Y., Kim, K. S. (2006). Genomic Comparison of Escherichia coli K1 Strains Isolated from the Cerebrospinal Fluid of Patients with Meningitis. Infect. Immun. 74: 2196-2206 [Abstract] [Full Text]  
• Honegger, A., Spinelli, S., Cambillau, C., Pluckthun, A. (2005). A mutation designed to alter crystal packing permits structural analysis of a tight-binding fluorescein-scFv complex. Protein Sci. 14: 2537-2549 [Abstract] [Full Text]  
• Reiling, S. A., Jansen, J. A., Henley, B. J., Singh, S., Chattin, C., Chandler, M., Rowen, D. W. (2005). Prc protease promotes mucoidy in mucA mutants of Pseudomonas aeruginosa. Microbiology 151: 2251-2261 [Abstract] [Full Text]  
• Paul, B. J., Berkmen, M. B., Gourse, R. L. (2005). From the Cover: DksA potentiates direct activation of amino acid promoters by ppGpp. Proc. Natl. Acad. Sci. USA 102: 7823-7828 [Abstract] [Full Text]  
• Zahnd, C., Spinelli, S., Luginbuhl, B., Amstutz, P., Cambillau, C., Pluckthun, A. (2004). Directed in Vitro Evolution and Crystallographic Analysis of a Peptide-binding Single Chain Antibody Fragment (scFv) with Low Picomolar Affinity. J. Biol. Chem. 279: 18870-18877 [Abstract] [Full Text]  
• Tomas, C. A., Beamish, J., Papoutsakis, E. T. (2004). Transcriptional Analysis of Butanol Stress and Tolerance in Clostridium acetobutylicum. J. Bacteriol. 186: 2006-2018 [Abstract] [Full Text]  
• Lyon, W. R., Caparon, M. G. (2004). Role for Serine Protease HtrA (DegP) of Streptococcus pyogenes in the Biogenesis of Virulence Factors SpeB and the Hemolysin Streptolysin S. Infect. Immun. 72: 1618-1625 [Abstract] [Full Text]  
• Sheehan, B. J., Bosse, J. T., Beddek, A. J., Rycroft, A. N., Kroll, J. S., Langford, P. R. (2003). Identification of Actinobacillus pleuropneumoniae Genes Important for Survival during Infection in Its Natural Host. Infect. Immun. 71: 3960-3970 [Abstract] [Full Text]  
• Di Paolo, C., Willuda, J., Kubetzko, S., Lauffer, I., Tschudi, D., Waibel, R., Pluckthun, A., Stahel, R. A., Zangemeister-Wittke, U. (2003). A Recombinant Immunotoxin Derived from a Humanized Epithelial Cell Adhesion Molecule-specific Single-Chain Antibody Fragment Has Potent and Selective Antitumor Activity. Clin. Cancer Res. 9: 2837-2848 [Abstract] [Full Text]  
• Huang, L., McCluskey, M. P., Ni, H., LaRossa, R. A. (2002). Global Gene Expression Profiles of the Cyanobacterium Synechocystis sp. Strain PCC 6803 in Response to Irradiation with UV-B and White Light. J. Bacteriol. 184: 6845-6858 [Abstract] [Full Text]  
• Spiers, A., Lamb, H. K., Cocklin, S., Wheeler, K. A., Budworth, J., Dodds, A. L., Pallen, M. J., Maskell, D. J., Charles, I. G., Hawkins, A. R. (2002). PDZ Domains Facilitate Binding of High Temperature Requirement Protease A (HtrA) and Tail-specific Protease (Tsp) to Heterologous Substrates through Recognition of the Small Stable RNA A (ssrA)-encoded Peptide. J. Biol. Chem. 277: 39443-39449 [Abstract] [Full Text]  
• Hengge-Aronis, R. (2002). Signal Transduction and Regulatory Mechanisms Involved in Control of the {sigma}S (RpoS) Subunit of RNA Polymerase. Microbiol. Mol. Biol. Rev. 66: 373-395 [Abstract] [Full Text]  
• Brown, L., Gentry, D., Elliott, T., Cashel, M. (2002). DksA Affects ppGpp Induction of RpoS at a Translational Level. J. Bacteriol. 184: 4455-4465 [Abstract] [Full Text]  
• Firoved, A. M., Boucher, J. C., Deretic, V. (2002). Global Genomic Analysis of AlgU ({sigma}E)-Dependent Promoters (Sigmulon) in Pseudomonas aeruginosa and Implications for Inflammatory Processes in Cystic Fibrosis. J. Bacteriol. 184: 1057-1064 [Abstract] [Full Text]  
• Innes, D., Beacham, I. R., Beven, C.-A., Douglas, M., Laird, M. W., Joly, J. C., Burns, D. M. (2001). The cryptic ushA gene (ushAc) in natural isolates of Salmonella enterica (serotype Typhimurium) has been inactivated by a single missense mutation. Microbiology 147: 1887-1896 [Abstract] [Full Text]  
• Branny, P., Pearson, J. P., Pesci, E. C., Köhler, T., Iglewski, B. H., Van Delden, C. (2001). Inhibition of Quorum Sensing by a Pseudomonas aeruginosa dksA Homologue. J. Bacteriol. 183: 1531-1539 [Abstract] [Full Text]  
• Bessette, P. H., Qiu, J., Bardwell, J. C. A., Swartz, J. R., Georgiou, G. (2001). Effect of Sequences of the Active-Site Dipeptides of DsbA and DsbC on In Vivo Folding of Multidisulfide Proteins in Escherichia coli. J. Bacteriol. 183: 980-988 [Abstract] [Full Text]  
• Schwesinger, F., Ros, R., Strunz, T., Anselmetti, D., Guntherodt, H.-J., Honegger, A., Jermutus, L., Tiefenauer, L., Pluckthun, A. (2000). Unbinding forces of single antibody-antigen complexes correlate with their thermal dissociation rates. Proc. Natl. Acad. Sci. USA 97: 9972-9977 [Abstract] [Full Text]  
• Willuda, J., Honegger, A., Waibel, R., Schubiger, P. A., Stahel, R., Zangemeister-Wittke, U., Pluckthun, A. (1999). High Thermal Stability Is Essential for Tumor Targeting of Antibody Fragments: Engineering of a Humanized Anti-epithelial Glycoprotein-2 (Epithelial Cell Adhesion Molecule) Single-Chain Fv Fragment. Cancer Res. 59: 5758-5767 [Abstract] [Full Text]  
• Ades, S. E., Connolly, L. E., Alba, B. M., Gross, C. A. (1999). The Escherichia coli sigma E-dependent extracytoplasmic stress response is controlled by the regulated proteolysis of an anti-sigma factor. Genes Dev. 13: 2449-2461 [Abstract] [Full Text]  
• Smeds, A., Varmanen, P., Palva, A. (1998). Molecular Characterization of a Stress-Inducible Gene from Lactobacillus helveticus. J. Bacteriol. 180: 6148-6153 [Abstract] [Full Text]  
• Kihara, A., Ito, K. (1998). Translocation, Folding, and Stability of the HflKC Complex with Signal Anchor Topogenic Sequences. J. Biol. Chem. 273: 29770-29775 [Abstract] [Full Text]  
• Berlyn, M. K. B. (1998). Linkage Map of Escherichia coli K-12, Edition 10: The Traditional Map. Microbiol. Mol. Biol. Rev. 62: 814-984 [Abstract] [Full Text]  
• Ros, R., Schwesinger, F., Anselmetti, D., Kubon, M., Schafer, R., Pluckthun, A., Tiefenauer, L. (1998). Antigen binding forces of individually addressed single-chain Fv antibody molecules. Proc. Natl. Acad. Sci. USA 95: 7402-7405 [Abstract] [Full Text]  
• Turner, A. K., Lovell, M. A., Hulme, S. D., Zhang-Barber, L., Barrow, P. A. (1998). Identification of Salmonella typhimurium Genes Required for Colonization of the Chicken Alimentary Tract and for Virulence in Newly Hatched Chicks. Infect. Immun. 66: 2099-2106 [Abstract] [Full Text]  
• Mohr, C. D., MacKichan, J. K., Shapiro, L. (1998). A Membrane-Associated Protein, FliX, Is Required for an Early Step in Caulobacter Flagellar Assembly. J. Bacteriol. 180: 2175-2185 [Abstract] [Full Text]  
• Joly, J. C., Leung, W. S., Swartz, J. R. (1998). Overexpression of Escherichia coli oxidoreductases increases recombinant insulin-like growth factor-I accumulation. Proc. Natl. Acad. Sci. USA 95: 2773-2777 [Abstract] [Full Text]  
• Loosmore, S. M., Yang, Y.-p., Oomen, R., Shortreed, J. M., Coleman, D. C., Klein, M. H. (1998). The Haemophilus influenzae HtrA Protein Is a Protective Antigen. Infect. Immun. 66: 899-906 [Abstract] [Full Text]  
• Gasc, A., Giammarinaro, P, Richter, S, Sicard, M (1998). Organization around the dnaA gene of Streptococcus pneumoniae. Microbiology 144: 433-439 [Abstract]  
• Skorko-Glonek, J., Skórko-Glonek, J., Krzewski, K., Zolese, G., Bertoli, E., Tanfani, F. (1997). HtrA Heat Shock Protease Interacts with Phospholipid Membranes and Undergoes Conformational Changes. J. Biol. Chem. 272: 8974-8982 [Abstract] [Full Text]  
• Willuda, J., Kubetzko, S., Waibel, R., Schubiger, P. A., Zangemeister-Wittke, U., Pluckthun, A. (2001). Tumor Targeting of Mono-, Di-, and Tetravalent Anti-p185HER-2 Miniantibodies Multimerized by Self-associating Peptides. J. Biol. Chem. 276: 14385-14392 [Abstract] [Full Text]