Analyses of Mlc-IIBGlc Interaction and a Plausible Molecular Mechanism of Mlc Inactivation by Membrane Sequestration

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2008 Mar 6

PMID 18319344

Citations 23

Authors

Tae-Wook Nam

Ha Il Jung

Young Jun An

Young-Ha Park

Sang Hee Lee

Yeong-Jae Seok

Sun-Shin Cha

Affiliations

Soon will be listed here.

Abstract

In Escherichia coli, glucose-dependent transcriptional induction of genes encoding a variety of sugar-metabolizing enzymes and transport systems is mediated by the phosphorylation state-dependent interaction of membrane-bound enzyme IICB(Glc) (EIICB(Glc)) with the global repressor Mlc. Here we report the crystal structure of a tetrameric Mlc in a complex with four molecules of enzyme IIB(Glc) (EIIB), the cytoplasmic domain of EIICB(Glc). Each monomer of Mlc has one bound EIIB molecule, indicating the 1:1 stoichiometry. The detailed view of the interface, along with the high-resolution structure of EIIB containing a sulfate ion at the phosphorylation site, suggests that the phosphorylation-induced steric hindrance and disturbance of polar intermolecular interactions impede complex formation. Furthermore, we reveal that Mlc possesses a built-in flexibility for the structural adaptation to its target DNA and that interaction of Mlc with EIIB fused only to dimeric proteins resulted in the loss of its DNA binding ability, suggesting that flexibility of the Mlc structure is indispensable for its DNA binding.

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References

Lee S, Boos W, Bouche J, Plumbridge J . Signal transduction between a membrane-bound transporter, PtsG, and a soluble transcription factor, Mlc, of Escherichia coli. EMBO J. 2000; 19(20):5353-61. PMC: 313994. DOI: 10.1093/emboj/19.20.5353. View

Kimata K, Inada T, Tagami H, Aiba H . A global repressor (Mlc) is involved in glucose induction of the ptsG gene encoding major glucose transporter in Escherichia coli. Mol Microbiol. 1998; 29(6):1509-19. DOI: 10.1046/j.1365-2958.1998.01035.x. View

Xia B, Vlamis-Gardikas A, Holmgren A, Wright P, Dyson H . Solution structure of Escherichia coli glutaredoxin-2 shows similarity to mammalian glutathione-S-transferases. J Mol Biol. 2001; 310(4):907-18. DOI: 10.1006/jmbi.2001.4721. View

Tanaka Y, Itoh F, Kimata K, Aiba H . Membrane localization itself but not binding to IICB is directly responsible for the inactivation of the global repressor Mlc in Escherichia coli. Mol Microbiol. 2004; 53(3):941-51. DOI: 10.1111/j.1365-2958.2004.04179.x. View

Lee C, Koo B, Cho S, Kim Y, Yoon M, Peterkofsky A . Requirement of the dephospho-form of enzyme IIANtr for derepression of Escherichia coli K-12 ilvBN expression. Mol Microbiol. 2005; 58(1):334-44. DOI: 10.1111/j.1365-2958.2005.04834.x. View

Nam T, Cho S, Shin D, Kim J, Jeong J, Lee J . The Escherichia coli glucose transporter enzyme IICB(Glc) recruits the global repressor Mlc. EMBO J. 2001; 20(3):491-8. PMC: 133465. DOI: 10.1093/emboj/20.3.491. View

Cai M, Williams Jr D, Wang G, Lee B, Peterkofsky A, Clore G . Solution structure of the phosphoryl transfer complex between the signal-transducing protein IIAGlucose and the cytoplasmic domain of the glucose transporter IICBGlucose of the Escherichia coli glucose phosphotransferase system. J Biol Chem. 2003; 278(27):25191-206. DOI: 10.1074/jbc.M302677200. View

Kim S, Nam T, Shin D, Koo B, Seok Y, Ryu S . Purification of Mlc and analysis of its effects on the pts expression in Escherichia coli. J Biol Chem. 1999; 274(36):25398-402. DOI: 10.1074/jbc.274.36.25398. View

Charpentier B, Bardey V, Robas N, Branlant C . The EIIGlc protein is involved in glucose-mediated activation of Escherichia coli gapA and gapB-pgk transcription. J Bacteriol. 1998; 180(24):6476-83. PMC: 107748. DOI: 10.1128/JB.180.24.6476-6483.1998. View

10.

Tanaka Y, Kimata K, Aiba H . A novel regulatory role of glucose transporter of Escherichia coli: membrane sequestration of a global repressor Mlc. EMBO J. 2000; 19(20):5344-52. PMC: 314007. DOI: 10.1093/emboj/19.20.5344. View

11.

Lee C, Cho S, Yoon M, Peterkofsky A, Seok Y . Escherichia coli enzyme IIANtr regulates the K+ transporter TrkA. Proc Natl Acad Sci U S A. 2007; 104(10):4124-9. PMC: 1794712. DOI: 10.1073/pnas.0609897104. View

12.

Jiang S, Tovchigrechko A, Vakser I . The role of geometric complementarity in secondary structure packing: a systematic docking study. Protein Sci. 2003; 12(8):1646-51. PMC: 2323951. DOI: 10.1110/ps.0304503. View

13.

Eberstadt M, Grdadolnik S, Gemmecker G, Kessler H, Buhr A, Erni B . Solution structure of the IIB domain of the glucose transporter of Escherichia coli. Biochemistry. 1996; 35(35):11286-92. DOI: 10.1021/bi960492l. View

14.

Plumbridge J . Control of the expression of the manXYZ operon in Escherichia coli: Mlc is a negative regulator of the mannose PTS. Mol Microbiol. 1998; 27(2):369-80. DOI: 10.1046/j.1365-2958.1998.00685.x. View

15.

Plumbridge J . Expression of the phosphotransferase system both mediates and is mediated by Mlc regulation in Escherichia coli. Mol Microbiol. 1999; 33(2):260-73. DOI: 10.1046/j.1365-2958.1999.01462.x. View

16.

Tanaka Y, Kimata K, Inada T, Tagami H, Aiba H . Negative regulation of the pts operon by Mlc: mechanism underlying glucose induction in Escherichia coli. Genes Cells. 1999; 4(7):391-9. DOI: 10.1046/j.1365-2443.1999.00268.x. View

17.

Park Y, Lee B, Seok Y, Peterkofsky A . In vitro reconstitution of catabolite repression in Escherichia coli. J Biol Chem. 2006; 281(10):6448-54. DOI: 10.1074/jbc.M512672200. View

18.

Plumbridge J . Expression of ptsG, the gene for the major glucose PTS transporter in Escherichia coli, is repressed by Mlc and induced by growth on glucose. Mol Microbiol. 1998; 29(4):1053-63. DOI: 10.1046/j.1365-2958.1998.00991.x. View

19.

Decker K, Plumbridge J, Boos W . Negative transcriptional regulation of a positive regulator: the expression of malT, encoding the transcriptional activator of the maltose regulon of Escherichia coli, is negatively controlled by Mlc. Mol Microbiol. 1998; 27(2):381-90. DOI: 10.1046/j.1365-2958.1998.00694.x. View

20.

Seitz S, Lee S, Pennetier C, Boos W, Plumbridge J . Analysis of the interaction between the global regulator Mlc and EIIBGlc of the glucose-specific phosphotransferase system in Escherichia coli. J Biol Chem. 2003; 278(12):10744-51. DOI: 10.1074/jbc.M212066200. View