Specialized roles of the two pathways for the synthesis of mannosylglycerate in osmoadaptation and thermoadaptation of Rhodothermus marinus

被引:41
作者
Borges, N
Marugg, JD
Empadinhas, N
da Costa, MS
Santos, H
机构
[1] Univ Nova Lisboa, Inst Tecnol Quim & Biol, P-2780156 Oeiras, Portugal
[2] Univ Coimbra, Dept Bioquim, P-3000 Coimbra, Portugal
[3] Univ Coimbra, Ctr Neurociencias & Biol Mol, P-3000 Coimbra, Portugal
[4] Nestle Res Ctr, CH-1000 Lausanne 26, Switzerland
关键词
D O I
10.1074/jbc.M312186200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Rhodothermus marinus responds to fluctuations in the growth temperature and/or salinity by accumulating mannosylglycerate (MG). Two alternative pathways for the synthesis of MG have been identified in this bacterium: a single-step pathway and a two-step pathway. In this work, the genetic and biochemical characterization of the two-step pathway was carried out with the goal of understanding the function of the two pathways and their regulatory mechanisms. Mannosyl-3-phosphoglycerate synthase (MPGS) of the two-step pathway was purified from R. marinus. Sequence information led to the isolation of two contiguous genes, mpgs (encoding MPGS) and mpgp ( encoding mannosyl-3-phosphoglycerate phosphatase). The recombinant MPGS had a low specific activity compared with other homologous MPGSs and contained similar to30 additional residues at the C terminus. Truncation of this extension produced a protein with a 10-fold higher specific activity. Moreover, the activity of the complete MPGS was enhanced upon incubation with R. marinus cell extracts, and protease inhibitors abolished activation. Therefore, the C-terminal peptide of MPGS was identified as a regulatory site for short term control of MG synthesis in R. marinus. The control of gene expression by heat and osmotic stress was also studied; the level of mannosylglycerate synthase involved in the single-step pathway was selectively enhanced by heat stress, whereas MPGS was overproduced in response to osmotic stress. The concomitant changes in the level of MG were assessed as well. We conclude that the two alternative pathways for the synthesis of MG are differently regulated at the level of expression to play specific roles in the adaptation of R. marinus to two different types of aggression. This is the only example of pathway multiplicity being rationalized in terms of the need to respond efficiently to distinct environmental stresses.
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页码:9892 / 9898
页数:7
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