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Molecular Pathogenesis

The Stress-Responsive dgk Gene from Streptococcus mutans Encodes a Putative Undecaprenol Kinase Activity

Maciej Lis, Howard K. Kuramitsu
Maciej Lis
Department of Oral Biology, State University of New York, Buffalo, New York 14214
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Howard K. Kuramitsu
Department of Oral Biology, State University of New York, Buffalo, New York 14214
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  • For correspondence: Kuramits@buffalo.edu
DOI: 10.1128/IAI.71.4.1938-1943.2003
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    FIG. 1.

    Membrane preparations of S. mutans strains GS-5 and Tn-1 carrying vector pTS749 (GS-5 vector and Tn-1 vector) and Tn-1 carrying plasmid pDGK749 (Tn-1 dgk) were assayed for DGK activity in the presence of lipid extracts from S. mutans. Error bars represent standard errors of the means for reactions performed in duplicate.

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    FIG. 2.

    DGK activities in membrane preparations of E. coli strain DH5α and DGK-deficient mutant RZ6 carrying vector pTrc99A and RZ6 expressing wild-type (wt) and mutated (mu) forms of dgk from S. mutans on plasmids pTrcDGK-1 and pTrcDGKTn-31, respectively. The reactions were performed in the presence of lipid extracts from S. mutans. Error bars represent standard errors of the means for reactions performed in duplicate.

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    Comparison of TLC separations of phospholipids generated by two different kinases. Lanes 1, 2, and 4, TLC separations of phospholipids generated by E. coli kinase from diacylglycerol (16:0), from S. mutans lipids, and from diacylglycerol (18:1), respectively; lane 3, separation of products of reactions catalyzed by membranes isolated from S. mutans and by purified DGK from E. coli by using S. mutans lipids as substrates. The products of both reactions were mixed prior to TLC separation.

  • FIG. 4.
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    FIG. 4.

    Phosphorylation of the lipid extract from S. mutans (L) or of undecaprenol (U). The sources of lipid kinase activities used were isolated membranes of S. mutans strains: GS-5 (G) or Tn-1 expressing S. mutans dgk on a plasmid (T/D) or purified DGK from E. coli (E).

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    FIG. 5.

    Saponification of phospholipids generated from undecaprenol by membranes isolated from S. mutans (S. m) or by purified DGK from E. coli (E. c).

  • FIG. 6.
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    FIG. 6.

    Generation of phospholipids from undecaprenol (U) and diacylglycerol (DG) by S. mutans membranes (S), by purified kinase from S. mutans (pS), and by purified DGK from E. coli (pE).

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The Stress-Responsive dgk Gene from Streptococcus mutans Encodes a Putative Undecaprenol Kinase Activity
Maciej Lis, Howard K. Kuramitsu
Infection and Immunity Apr 2003, 71 (4) 1938-1943; DOI: 10.1128/IAI.71.4.1938-1943.2003

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The Stress-Responsive dgk Gene from Streptococcus mutans Encodes a Putative Undecaprenol Kinase Activity
Maciej Lis, Howard K. Kuramitsu
Infection and Immunity Apr 2003, 71 (4) 1938-1943; DOI: 10.1128/IAI.71.4.1938-1943.2003
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KEYWORDS

Heat-Shock Response
Phosphotransferases (Alcohol Group Acceptor)
Streptococcus mutans

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