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Solution structures of the inactive and BeF3-activated response regulator CheY2.

Riepl H, Scharf B, Schmitt R, Kalbitzer HR, Maurer T
J Mol Biol 338 287-97 (2004)
Cited: 11 times
EuropePMC logo PMID: 15066432

Abstract

The chemotactic signalling chain to the flagellar motor of Sinorhizobium meliloti features a new type of response regulator, CheY2. CheY2 activated by phosphorylation (CheY2-P) controls the rotary speed of the flagellar motor (instead of reversing the sense of rotation), and it is efficiently dephosphorylated by phospho-retrotransfer to the cognate kinase, CheA. Here, we report the NMR solution structures of the Mg(2+)-complex of inactive CheY2, and of activated CheY2-BeF(3), a stable analogue of CheY2-P, to an overall root mean square deviation of 0.042 nm and 0.027 nm, respectively. The 14 kDa CheY2 protein exhibits a characteristic open (alpha/beta)(5) conformation. Modification of CheY2 by BeF(3)(-) leads to large conformational changes of the protein, which are in the limits of error identical with those observed by phosphorylation of the active-centre residue Asp58. In BeF(3)-activated CheY2, the position of Thr88-OH favours the formation of a hydrogen bond with the active site, Asp58-BeF(3), similar to BeF(3)-activated CheY from Escherichia coli. In contrast to E.coli, this reorientation is not involved in a Tyr-Thr-coupling mechanism, that propagates the signal from the incoming phosphoryl group to the C-terminally located FliM-binding surface. Rather, a rearrangement of the Phe59 side-chain to interact with Ile86-Leu95-Val96 along with a displacement of alpha4 towards beta5 is stabilised in S.meliloti. The resulting, activation-induced, compact alpha4-beta5-alpha5 surface forms a unique binding domain suited for specific interaction with and signalling to a rotary motor that requires a gradual speed control. We propose that these new features of response regulator activation, compared to other two-component systems, are the key for the observed unique phosphorylation, dephosphorylation and motor control mechanisms in S.meliloti.

Articles - 1p6u mentioned but not cited (2)

  1. Crystal structure of activated CheY1 from Helicobacter pylori. Lam KH, Ling TK, Au SW. J Bacteriol 192 2324-2334 (2010)
  2. Expression, purification and preliminary crystallographic analysis of Pseudomonas aeruginosa RocR protein. Kotaka M, Dutta S, Lee HC, Lim MJ, Wong Y, Rao F, Mitchell EP, Liang ZX, Lescar J. Acta Crystallogr Sect F Struct Biol Cryst Commun 65 1035-1038 (2009)


Articles citing this publication (9)

  1. The CheYs of Rhodobacter sphaeroides. Porter SL, Wadhams GH, Martin AC, Byles ED, Lancaster DE, Armitage JP. J Biol Chem 281 32694-32704 (2006)
  2. Crystal structures of the response regulator DosR from Mycobacterium tuberculosis suggest a helix rearrangement mechanism for phosphorylation activation. Wisedchaisri G, Wu M, Sherman DR, Hol WG. J Mol Biol 378 227-242 (2008)
  3. Control of speed modulation (chemokinesis) in the unidirectional rotary motor of Sinorhizobium meliloti. Attmannspacher U, Scharf B, Schmitt R. Mol Microbiol 56 708-718 (2005)
  4. Insights into correlated motions and long-range interactions in CheY derived from molecular dynamics simulations. Knaggs MH, Salsbury FR, Edgell MH, Fetrow JS. Biophys J 92 2062-2079 (2007)
  5. Interaction of CheY2 and CheY2-P with the cognate CheA kinase in the chemosensory-signalling chain of Sinorhizobium meliloti. Riepl H, Maurer T, Kalbitzer HR, Meier VM, Haslbeck M, Schmitt R, Scharf B. Mol Microbiol 69 1373-1384 (2008)
  6. Nuclear magnetic resonance structure and dynamics of the response regulator Sma0114 from Sinorhizobium meliloti. Sheftic SR, Garcia PP, White E, Robinson VL, Gage DJ, Alexandrescu AT. Biochemistry 51 6932-6941 (2012)
  7. Phosphorylation-induced activation of the response regulator VraR from Staphylococcus aureus: insights from hydrogen exchange mass spectrometry. Liu YH, Belcheva A, Konermann L, Golemi-Kotra D. J Mol Biol 391 149-163 (2009)
  8. Phospho-dependent signaling during the general stress response by the atypical response regulator and ClpXP adaptor RssB. Schwartz J, Son J, Brugger C, Deaconescu AM. Protein Sci 30 899-907 (2021)
  9. (1)H, (13)C and (15)N resonance assignments for the response regulator CheY3 from Rhodobacter sphaeroides. Varela L, Bell CH, Armitage JP, Redfield C. Biomol NMR Assign 10 373-378 (2016)


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