Crystal structure of a phosphatase-resistant mutant of sporulation response regulator Spo0F from Bacillus subtilis

Madhusudan, M. D.; Zapf, James; Whiteley, John M.; Hoch, James A.; Xuong, N.-H.; Varughese, Kottayil I.

doi:10.1016/s0969-2126(96)00074-3

Cited by 78 publications

(90 citation statements)

References 55 publications

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“…One of the water molecules was hydrogen bonded to the main chain carbonyl oxygen of Gln-55. The coordination of the metal to the third acidic residue (Glu-9) and the absence of direct coordination to the main chain carbonyl oxygen of Gln-55 differs from what has been reported for all metal-bound receiver domains (32)(33)(34)(35)(36).…”

Section: Crystal Structure Of the Response Regulator Divkcontrasting

confidence: 69%

Crystallographic and Biochemical Studies of DivK Reveal Novel Features of an Essential Response Regulator in Caulobacter crescentus

Guillet¹,

Ohta²,

Cabantous³

et al. 2002

Journal of Biological Chemistry

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Section: Crystal Structure Of the Response Regulator Divkcontrasting

confidence: 69%

Crystallographic and Biochemical Studies of DivK Reveal Novel Features of an Essential Response Regulator in Caulobacter crescentus

Guillet¹,

Ohta²,

Cabantous³

et al. 2002

Journal of Biological Chemistry

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“…YneI is most probably a single-domain response regulator. The sequence of this 120-residue protein is very similar to that of CheY and Spo0F, for which the three-dimensional structures are known (22,40). YneJ is a predicted 160-residue integral membrane protein without clear similarity to any protein sequence available in the databases.…”

mentioning

confidence: 92%

Efficient Spore Synthesis in Bacillus subtilis Depends on the CcdA Protein

Schiött

Hederstedt

2000

J Bacteriol

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CcdA is known to be required for the synthesis of c-type cytochromes in Bacillus subtilis, but the exact function of this membrane protein is not known. We show that CcdA also plays a role in spore synthesis. The expression of ccdA and the two downstream genes yneI and yneJ was analyzed. There is a promoter for each gene, but there is only one transcription terminator, located after the yneJ gene. The promoter for ccdA was found to be weak and was active mainly during the transition from exponential growth to stationary phase. The promoters for yneI and yneJ were both active in the exponential growth phase. The levels of the CcdA and YneJ proteins in the membrane were consistent with the observed promoter activities. The ccdA promoter activity was independent of whether the ccdA-yneI-yneJ gene products were absent or overproduced in the cell. It is shown that the four known cytochromes c in B. subtilis and the YneI and YneJ proteins are not required for sporulation. The combined data from analysis of sporulation-specific sigma factor activity, resistance properties of spores, and spore morphology indicate that CcdA deficiency affects stage V in sporulation. We conclude that CcdA, YneI, and YneJ are functionally unrelated proteins and that the role of CcdA in cytochrome c and spore synthesis probably relates to sulfhydryl redox chemistry on the outer surface of the cytoplasmic membrane.

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“…This section focuses on the crystallographic results, and NMR studies are discussed in a different section. Structural analysis has been carried out on the metalfree form (17) and on the metal-bound forms (18,19). Spo0F, like all other response regulators, has an ␣/␤ fold with a central ␤-sheet and five helices surrounding it (Fig.…”

Section: Structural Studies Of Spo0fmentioning

confidence: 99%

Conformational Changes of Spo0F along the Phosphotransfer Pathway

Varughese

2005

J Bacteriol

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Spo0F is a secondary messenger in the sporulation phosphorelay, and its structure has been characterized crystallographically in the apo-state, in the metal-bound state, and in an interacting state with a phosphotransferase. Additionally, the solution structure of the molecule has been characterized by nuclear magnetic resonance techniques in the unliganded state and in complex with beryllofluoride. Spo0F is a single-domain protein with a well-defined three-dimensional structure, but it is capable of adapting to specific conformations for catching and releasing the phosphoryl moiety. This commentary deals with the conformational fluctuations of the molecule as it moves from an apo-state to a metal-coordinated state, to a phosphorylated state, and then to a phosphoryl-transferring state.Spo0F is a component of the sporulation phosphorelay of Bacillus subtilis (10) which acts as a gateway for all sporulation signals processed by various histidine kinases (Fig. 1). The initiation of sporulation is controlled by a single parameter, the degree of phosphorylation of the transcription factor Spo0A. It is Spo0F that takes the phosphoryl moiety from the sensor kinases and passes it on to Spo0A through the phosphotransferase Spo0B. Hence, the activity of Spo0F involves interactions with the sensor kinases and Spo0B in receiving and donating the phosphoryl group. Therefore, to describe the conformational pathway of Spo0F, it is necessary to consider four distinct conformational states for Spo0F: (i) isolated unphosphorylated state, (ii) unphosphorylated Spo0F interacting with KinA, (iii) phosphorylated Spo0F, and (iv) phosphorylated Spo0F interacting with Spo0B. Although the documented structural data are inadequate to fully describe all these distinct states, the available data are reviewed here to gain the best understanding of the conformational states of Spo0F. STRUCTURAL STUDIES OF Spo0FUnphosphorylated state. The structure of Spo0F has been studied by both crystallographic and nuclear magnetic resonance (NMR) techniques. This section focuses on the crystallographic results, and NMR studies are discussed in a different section. Structural analysis has been carried out on the metalfree form (17) and on the metal-bound forms (18,19). Spo0F, like all other response regulators, has an ␣/␤ fold with a central ␤-sheet and five helices surrounding it (Fig. 2). The active site is located on the top of the ␤-strands and comprises an acid pocket with three conserved aspartates: Asp10, Asp11, and Asp54. The site of phosphorylation, Asp54, is located at the bottom of a shallow pocket. All phosphoryl transfer reactions make use of cations, and in the two component/phosphorelay systems, it is the response regulators that bind the cations and present them for catalysis. The crystal structures of the metalfree and metal-bound forms are very similar, and the changes are confined mainly to the metal binding site. The most significant difference between the two states of Spo0F is that, in the metal-free structure, the metal cavity is no...

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Crystal structure of a phosphatase-resistant mutant of sporulation response regulator Spo0F from Bacillus subtilis

Cited by 78 publications

References 55 publications

Crystallographic and Biochemical Studies of DivK Reveal Novel Features of an Essential Response Regulator in Caulobacter crescentus

Crystallographic and Biochemical Studies of DivK Reveal Novel Features of an Essential Response Regulator in Caulobacter crescentus

Efficient Spore Synthesis in Bacillus subtilis Depends on the CcdA Protein

Conformational Changes of Spo0F along the Phosphotransfer Pathway

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