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. 1993 Oct;175(20):6546–6552. doi: 10.1128/jb.175.20.6546-6552.1993

Functional exchangeability of the ABC proteins of the periplasmic binding protein-dependent transport systems Ugp and Mal of Escherichia coli.

D Hekstra 1, J Tommassen 1
PMCID: PMC206765  PMID: 8407831

Abstract

The periplasmic binding protein-dependent transport systems Ugp and Mal of Escherichia coli transport sn-glycerol-3-phosphate and maltose, respectively. The UgpC and MalK proteins of these transport systems, which couple energy to the transport process by ATP-hydrolysis, are highly homologous, suggesting that they might be functionally exchangeable. Complementation experiments showed that UgpC expression could restore growth of a malK mutant on maltose as a carbon source, provided that it was expressed at a sufficiently high level in the absence of the integral inner membrane components UgpA and/or UgpE of the Ugp system. Conversely, MalK expression could complement ugpC mutants and restore the utilization of sn-glycerol-3-phosphate as a phosphate source. The hybrid transporters appeared to be less efficient than the wild-type systems. The complementation of ugpC mutations by MalK was strongly inhibited by the presence of glucose or alpha-methylglucoside, which are substrates of the phosphotransferase system. This inhibition is probably due to hypersensitivity of the hybrid UgpBAE-MalK transporter to inducer exclusion. UgpC expression did not complement the regulatory function of MalK in mal gene expression. The exchangeability of UgpC and MalK indicates that these proteins do not contribute to a substrate-binding site conferring substrate specificity to the transporter. These are the first examples of functional, hybrid periplasmic permeases in which the energy-coupling components could be functionally exchanged.

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Selected References

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  1. Ambudkar S. V., Larson T. J., Maloney P. C. Reconstitution of sugar phosphate transport systems of Escherichia coli. J Biol Chem. 1986 Jul 15;261(20):9083–9086. [PubMed] [Google Scholar]
  2. Ames G. F., Joshi A. K. Energy coupling in bacterial periplasmic permeases. J Bacteriol. 1990 Aug;172(8):4133–4137. doi: 10.1128/jb.172.8.4133-4137.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ames G. F., Mimura C. S., Shyamala V. Bacterial periplasmic permeases belong to a family of transport proteins operating from Escherichia coli to human: Traffic ATPases. FEMS Microbiol Rev. 1990 Aug;6(4):429–446. doi: 10.1111/j.1574-6968.1990.tb04110.x. [DOI] [PubMed] [Google Scholar]
  4. Argast M., Boos W. Co-regulation in Escherichia coli of a novel transport system for sn-glycerol-3-phosphate and outer membrane protein Ic (e, E) with alkaline phosphatase and phosphate-binding protein. J Bacteriol. 1980 Jul;143(1):142–150. doi: 10.1128/jb.143.1.142-150.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bremer E., Silhavy T. J., Weinstock G. M. Transposable lambda placMu bacteriophages for creating lacZ operon fusions and kanamycin resistance insertions in Escherichia coli. J Bacteriol. 1985 Jun;162(3):1092–1099. doi: 10.1128/jb.162.3.1092-1099.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brzoska P., Boos W. Characteristics of a ugp-encoded and phoB-dependent glycerophosphoryl diester phosphodiesterase which is physically dependent on the ugp transport system of Escherichia coli. J Bacteriol. 1988 Sep;170(9):4125–4135. doi: 10.1128/jb.170.9.4125-4135.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brzoska P., Boos W. The ugp-encoded glycerophosphoryldiester phosphodiesterase, a transport-related enzyme of Escherichia coli. FEMS Microbiol Rev. 1989 Jun;5(1-2):115–124. doi: 10.1016/0168-6445(89)90015-6. [DOI] [PubMed] [Google Scholar]
  8. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  9. Cozzarelli N. R., Freedberg W. B., Lin E. C. Genetic control of L-alpha-glycerophosphate system in Escherichia coli. J Mol Biol. 1968 Feb 14;31(3):371–387. doi: 10.1016/0022-2836(68)90415-4. [DOI] [PubMed] [Google Scholar]
  10. Davidson A. L., Nikaido H. Overproduction, solubilization, and reconstitution of the maltose transport system from Escherichia coli. J Biol Chem. 1990 Mar 15;265(8):4254–4260. [PubMed] [Google Scholar]
  11. Davidson A. L., Nikaido H. Purification and characterization of the membrane-associated components of the maltose transport system from Escherichia coli. J Biol Chem. 1991 May 15;266(14):8946–8951. [PubMed] [Google Scholar]
  12. Dean D. A., Reizer J., Nikaido H., Saier M. H., Jr Regulation of the maltose transport system of Escherichia coli by the glucose-specific enzyme III of the phosphoenolpyruvate-sugar phosphotransferase system. Characterization of inducer exclusion-resistant mutants and reconstitution of inducer exclusion in proteoliposomes. J Biol Chem. 1990 Dec 5;265(34):21005–21010. [PubMed] [Google Scholar]
  13. Ehrmann M., Boos W. Identification of endogenous inducers of the mal regulon in Escherichia coli. J Bacteriol. 1987 Aug;169(8):3539–3545. doi: 10.1128/jb.169.8.3539-3545.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Elish M. E., Pierce J. R., Earhart C. F. Biochemical analysis of spontaneous fepA mutants of Escherichia coli. J Gen Microbiol. 1988 May;134(5):1355–1364. doi: 10.1099/00221287-134-5-1355. [DOI] [PubMed] [Google Scholar]
  15. Elvin C. M., Hardy C. M., Rosenberg H. Pi exchange mediated by the GlpT-dependent sn-glycerol-3-phosphate transport system in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1054–1058. doi: 10.1128/jb.161.3.1054-1058.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ezaki B., Mori H., Ogura T., Hiraga S. Possible involvement of the ugpA gene product in the stable maintenance of mini-F plasmid in Escherichia coli. Mol Gen Genet. 1990 Sep;223(3):361–368. doi: 10.1007/BF00264441. [DOI] [PubMed] [Google Scholar]
  17. Gallagher M. P., Pearce S. R., Higgins C. F. Identification and localization of the membrane-associated, ATP-binding subunit of the oligopeptide permease of Salmonella typhimurium. Eur J Biochem. 1989 Mar 1;180(1):133–141. doi: 10.1111/j.1432-1033.1989.tb14623.x. [DOI] [PubMed] [Google Scholar]
  18. Higgins C. F., Hyde S. C., Mimmack M. M., Gileadi U., Gill D. R., Gallagher M. P. Binding protein-dependent transport systems. J Bioenerg Biomembr. 1990 Aug;22(4):571–592. doi: 10.1007/BF00762962. [DOI] [PubMed] [Google Scholar]
  19. Hoekstra W. P., Bergmans J. E., Zuidweg E. M. Role of recBC nuclease in Escherichia coli transformation. J Bacteriol. 1980 Aug;143(2):1031–1032. doi: 10.1128/jb.143.2.1031-1032.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hofnung M., Hatfield D., Schwartz M. malB region in Escherichia coli K-12: characterization of new mutations. J Bacteriol. 1974 Jan;117(1):40–47. doi: 10.1128/jb.117.1.40-47.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hyde S. C., Emsley P., Hartshorn M. J., Mimmack M. M., Gileadi U., Pearce S. R., Gallagher M. P., Gill D. R., Hubbard R. E., Higgins C. F. Structural model of ATP-binding proteins associated with cystic fibrosis, multidrug resistance and bacterial transport. Nature. 1990 Jul 26;346(6282):362–365. doi: 10.1038/346362a0. [DOI] [PubMed] [Google Scholar]
  22. Kasahara M., Makino K., Amemura M., Nakata A. Nucleotide sequence of the ugpQ gene encoding glycerophosphoryl diester phosphodiesterase of Escherichia coli K-12. Nucleic Acids Res. 1989 Apr 11;17(7):2854–2854. doi: 10.1093/nar/17.7.2854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kasahara M., Makino K., Amemura M., Nakata A., Shinagawa H. Dual regulation of the ugp operon by phosphate and carbon starvation at two interspaced promoters. J Bacteriol. 1991 Jan;173(2):549–558. doi: 10.1128/jb.173.2.549-558.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kerppola R. E., Shyamala V. K., Klebba P., Ames G. F. The membrane-bound proteins of periplasmic permeases form a complex. Identification of the histidine permease HisQMP complex. J Biol Chem. 1991 May 25;266(15):9857–9865. [PubMed] [Google Scholar]
  25. Kleerebezem M., Tommassen J. Expression of the pspA gene stimulates efficient protein export in Escherichia coli. Mol Microbiol. 1993 Mar;7(6):947–956. doi: 10.1111/j.1365-2958.1993.tb01186.x. [DOI] [PubMed] [Google Scholar]
  26. Kühnau S., Reyes M., Sievertsen A., Shuman H. A., Boos W. The activities of the Escherichia coli MalK protein in maltose transport, regulation, and inducer exclusion can be separated by mutations. J Bacteriol. 1991 Apr;173(7):2180–2186. doi: 10.1128/jb.173.7.2180-2186.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lerner C. G., Inouye M. Low copy number plasmids for regulated low-level expression of cloned genes in Escherichia coli with blue/white insert screening capability. Nucleic Acids Res. 1990 Aug 11;18(15):4631–4631. doi: 10.1093/nar/18.15.4631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  29. Lugtenberg B., Peters R., Bernheimer H., Berendsen W. Influence of cultural conditions and mutations on the composition of the outer membrane proteins of Escherichia coli. Mol Gen Genet. 1976 Sep 23;147(3):251–262. doi: 10.1007/BF00582876. [DOI] [PubMed] [Google Scholar]
  30. Meadow N. D., Fox D. K., Roseman S. The bacterial phosphoenolpyruvate: glycose phosphotransferase system. Annu Rev Biochem. 1990;59:497–542. doi: 10.1146/annurev.bi.59.070190.002433. [DOI] [PubMed] [Google Scholar]
  31. Mimura C. S., Holbrook S. R., Ames G. F. Structural model of the nucleotide-binding conserved component of periplasmic permeases. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):84–88. doi: 10.1073/pnas.88.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Overduin P., Boos W., Tommassen J. Nucleotide sequence of the ugp genes of Escherichia coli K-12: homology to the maltose system. Mol Microbiol. 1988 Nov;2(6):767–775. doi: 10.1111/j.1365-2958.1988.tb00088.x. [DOI] [PubMed] [Google Scholar]
  33. Postma P. W., Epstein W., Schuitema A. R., Nelson S. O. Interaction between IIIGlc of the phosphoenolpyruvate:sugar phosphotransferase system and glycerol kinase of Salmonella typhimurium. J Bacteriol. 1984 Apr;158(1):351–353. doi: 10.1128/jb.158.1.351-353.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Reyes M., Shuman H. A. Overproduction of MalK protein prevents expression of the Escherichia coli mal regulon. J Bacteriol. 1988 Oct;170(10):4598–4602. doi: 10.1128/jb.170.10.4598-4602.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Richarme G., el Yaagoubi A., Kohiyama M. The MglA component of the binding protein-dependent galactose transport system of Salmonella typhimurium is a galactose-stimulated ATPase. J Biol Chem. 1993 May 5;268(13):9473–9477. [PubMed] [Google Scholar]
  36. Saier M. H., Jr Protein phosphorylation and allosteric control of inducer exclusion and catabolite repression by the bacterial phosphoenolpyruvate: sugar phosphotransferase system. Microbiol Rev. 1989 Mar;53(1):109–120. doi: 10.1128/mr.53.1.109-120.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schneider E., Walter C. A chimeric nucleotide-binding protein, encoded by a hisP-malK hybrid gene, is functional in maltose transport in Salmonella typhimurium. Mol Microbiol. 1991 Jun;5(6):1375–1383. doi: 10.1111/j.1365-2958.1991.tb00784.x. [DOI] [PubMed] [Google Scholar]
  38. Schweizer H., Boos W. Characterization of the ugp region containing the genes for the phoB dependent sn-glycerol-3-phosphate transport system of Escherichia coli. Mol Gen Genet. 1984;197(1):161–168. doi: 10.1007/BF00327937. [DOI] [PubMed] [Google Scholar]
  39. Speiser D. M., Ames G. F. Salmonella typhimurium histidine periplasmic permease mutations that allow transport in the absence of histidine-binding proteins. J Bacteriol. 1991 Feb;173(4):1444–1451. doi: 10.1128/jb.173.4.1444-1451.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stewart G. S., Lubinsky-Mink S., Jackson C. G., Cassel A., Kuhn J. pHG165: a pBR322 copy number derivative of pUC8 for cloning and expression. Plasmid. 1986 May;15(3):172–181. doi: 10.1016/0147-619x(86)90035-1. [DOI] [PubMed] [Google Scholar]
  41. Su T. Z., Schweizer H. P., Oxender D. L. Carbon-starvation induction of the ugp operon, encoding the binding protein-dependent sn-glycerol-3-phosphate transport system in Escherichia coli. Mol Gen Genet. 1991 Nov;230(1-2):28–32. doi: 10.1007/BF00290646. [DOI] [PubMed] [Google Scholar]
  42. Tommassen J., Eiglmeier K., Cole S. T., Overduin P., Larson T. J., Boos W. Characterization of two genes, glpQ and ugpQ, encoding glycerophosphoryl diester phosphodiesterases of Escherichia coli. Mol Gen Genet. 1991 Apr;226(1-2):321–327. doi: 10.1007/BF00273621. [DOI] [PubMed] [Google Scholar]
  43. Tommassen J., Lugtenberg B. Outer membrane protein e of Escherichia coli K-12 is co-regulated with alkaline phosphatase. J Bacteriol. 1980 Jul;143(1):151–157. doi: 10.1128/jb.143.1.151-157.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  45. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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