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Structural Changes and Cellular Localization of Resuscitation-Promoting Factor in Environmental Isolates of Micrococcus luteus

  • Environmental Microbiology
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Abstract

Dormancy among nonsporulating actinobacteria is now a widely accepted phenomenon. In Micrococcus luteus, the resuscitation of dormant cells is caused by a small secreted protein (resuscitation-promoting factor, or Rpf) that is found in “spent culture medium.” Rpf is encoded by a single essential gene in M. luteus. Homologs of Rpf are widespread among the high G + C Gram-positive bacteria, including mycobacteria and streptomycetes, and most organisms make several functionally redundant proteins. M. luteus Rpf comprises a lysozyme-like domain that is necessary and sufficient for activity connected through a short linker region to a LysM motif, which is present in a number of cell-wall-associated enzymes. Muralytic activity is responsible for resuscitation. In this report, we characterized a number of environmental isolates of M. luteus, including several recovered from amber. There was substantial variation in the predicted rpf gene product. While the lysozyme-like and LysM domains showed little variation, the linker region was elongated from ten amino acid residues in the laboratory strains to as many as 120 residues in one isolate. The genes encoding these Rpf proteins have been characterized, and a possible role for the Rpf linker in environmental adaptation is proposed. The environmental isolates show enhanced resistance to lysozyme as compared with the laboratory strains and this correlates with increased peptidoglycan acetylation. In strains that make a protein with an elongated linker, Rpf was bound to the cell wall, rather than being released to the growth medium, as occurs in reference strains. This rpf gene was introduced into a lysozyme-sensitive reference strain. Both rpf genes were expressed in transformants which showed a slight but statistically significant increase in lysozyme resistance.

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Acknowledgements

The authors are grateful to the Center for the Study of Emerging Diseases and the “Program for Molecular and Cellular Biology” of the Russian Academy of Science for their generous financial support. We thank Maria Ines Zylber for helping with the figures.

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Authors and Affiliations

  1. Kuvin Centre and Department of Microbiology and Molecular Genetics, Hebrew University, P.O. Box 12272, Jerusalem, 91120, Israel

    Viktoria Koltunov & Charles L. Greenblatt

  2. Bakh Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, 117071, Moscow, Russia

    Anna V. Goncharenko, Galya R. Demina & Arseny S. Kaprelyants

  3. Department of Neuroscience, Columbia University, 1051 Riverside Drive, New York, NY, 10032, USA

    Benjamin Y. Klein

  4. Institute of Biological Environmental and Rural Sciences, University of Wales, Aberystwyth, Ceredigion, SY23 3DD, UK

    Michael Young

Authors
  1. Viktoria Koltunov
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  2. Charles L. Greenblatt
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  3. Anna V. Goncharenko
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  4. Galya R. Demina
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  5. Benjamin Y. Klein
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  6. Michael Young
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  7. Arseny S. Kaprelyants
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Correspondence to Charles L. Greenblatt.

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Koltunov, V., Greenblatt, C.L., Goncharenko, A.V. et al. Structural Changes and Cellular Localization of Resuscitation-Promoting Factor in Environmental Isolates of Micrococcus luteus . Microb Ecol 59, 296–310 (2010). https://doi.org/10.1007/s00248-009-9573-1

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  • DOI: https://doi.org/10.1007/s00248-009-9573-1

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