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Identification of nolR ‐regulated proteins in Sinorhizobium meliloti using proteome analysis
Author(s) -
Chen Hancai,
Higgins Jody,
Kondorosi Eva,
Kondorosi Adam,
Djordjevic Michael A.,
Weinman Jeremy J.,
Rolfe Barry G.
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200011)21:17<3823::aid-elps3823>3.0.co;2-k
Subject(s) - proteome , biochemistry , heat shock protein , biology , two dimensional gel electrophoresis , mutant , gel electrophoresis , proteomics , sinorhizobium meliloti , hsp60 , coomassie brilliant blue , microbiology and biotechnology , gene , staining , hsp70 , genetics
Abstract Extractable proteins from Sinorhizobium meliloti strains AK631 and EK698 (a Tn5‐induced nolR ‐deficient mutant of AK631), grown in tryptone agar (TA) medium with or without the addition of the plant signal luteolin, were separated by two‐dimensional gel electrophoresis and compared. Analysis of silver‐stained gels showed that the nolR mutant had 189 proteins that were significantly altered in their levels (101 protein spots up‐ and 88 downregulated). Coomassie‐stained preparative two‐dimensional (2‐D) gels or polyvinylidene difluoride (PVDF) membranes blotted from preparative gels showed that at least 52 of the altered proteins could be reproducibly detected and isolated from the nolR mutant. These 52 altered protein spots were classified into five groups based on an assessment of protein abundance by computer analysis and the effect of the presence or absence of luteolin addition to the growth medium. N ‐terminal microsequencing of 38 proteins revealed that the most striking feature of the consequence of the nolR mutation is the number and broad spectrum of cellular functions that are affected by the loss of the NolR function. These include proteins involved in the tricarboxylic acid (TCA) cycle, heat shock and cold shock proteins, protein synthesis, a translation elongation factor, oxidative stress and cell growth and maintenance functions. We propose that the NolR repressor is a global regulatory protein which responds to environmental factors to fine‐tune intracellular metabolism.