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Journal of Biochemistry Advance Access published online on June 1, 2007
Journal of Biochemistry, doi:10.1093/jb/mvm121
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© 2007 The Japanese Biochemical Society

Model based definition of population heterogeneity and its effects on metabolism in sporulating Bacillus subtilis

Mineo Morohashi1, Yoshiaki Ohashi1,2,*, Saeka Tani2, Kotaro Ishii2,{dagger}, Mitsuhiro Itaya2, Hideaki Nanamiya3,{ddagger}, Fujio Kawamura3, Masaru Tomita1,2 and Tomoyoshi Soga1,2

1 Human Metabolome Technologies, Inc., Tsuruoka, Yamagata 997-0052, Japan
2 Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata 997-0017, Japan
3 College of Science, Rikkyo (St. Paul's) University, Toshima-ku, Tokyo 171-8501, Japan

*Corresponding author: 246-2 Mizukami Kakuganji, Tsuruoka, Yamagata 997-0052, Japan. Tel: +81-235-25-1447, Fax: +81-235-25-1450, E-mail: ohashi{at}humanmetabolome.com / ohashi{at}sfc.keio.ac.jp

Received April 1, 2007; Accepted May 11, 2007


  Abstract

The soil bacterium Bacillus subtilis forms dormant, robust spores as a tactic to ensure survival under conditions of starvation. However, the sporulating culture includes sporulating and non-sporulating cells, because a portion of the cell population initiates sporulation in wild-type strain. We anticipated that the population effect must be considered carefully when to analyze samples yielding population heterogeneity. We first built a mathematical model and simulated for signal transduction of the sporulation cue to see what mechanisms are responsible for generating the heterogeneity. The simulated results were confirmed experimentally, where heterogeneity is primarily modulated by negative feedback circuits, resulting in generation of a bistable response within the sporulating culture. We also confirmed that mutants relevant to negative feedback yield either sporulating or non-sporulating subpopulations. To see the effect of molecular mechanism between sporulating and non-sporulating cells in distinct manner, metabolome analysis was conducted using the above mutants. The metabolic profiles exhibited distinct characteristics with time regardless of whether sporulation was initiated or not. In addition, several distinct characteristics of metabolites were observed between strains, which was inconsistent with previously reported data. The results imply that careful consideration must be made in the interpretation of data obtained from cells yielding population heterogeneity.

Key Words: heterogeneity, Bacillus subtilis, metabolome, sporulation

{dagger} Present address, The University of Tokyo, Kashiwa, Chiba 277-0000, Japan

{ddagger} Present address, Ehime University, Matsuyama, Ehime 790-8577, Japan


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