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Current Microbiology

, Volume 53, Issue 4, pp 311–316 | Cite as

Proteolytic Cleavage of a Spectrin-Related Protein by Calcium-Dependent Protease in Neurospora crassa

  • M. Cotado-Sampayo
  • M. Ojha
  • R. Ortega-Pérez
  • M.-L. Chappuis
  • F. BarjaEmail author
Article

Abstract

To investigate the functional significance of a cytoskeletal spectrin-like protein, we studied its localization pattern in Neurospora crassa and sought the answer to whether it is a substrate for another apically localized protein, the calcium-dependent protease (CDP II). Immunoblots of crude extracts from exponentially growing mycelia, separated by one- and two-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis using antichicken α/β-spectrin antibodies, revealed a single band of approximately relative mass (Mr) 100 kDa with an isoeletric point (pI) in the range of 6.5 to 7.0. Despite rigorous efforts, we could not confirm the presence of an Mr 240- to 220-kDa spectrin-like protein in N. crassa. The immunofluorescence- and immunogold-labeling Mr 100-kDa protein showed its predominance along the plasma membrane of the conidia during the swelling phase of germination. In contrast, in the germ tubes and the growing hyphae, the localization was polarized and concentrated mainly in the apical region. The in vitro proteolysis experiments showed that indeed this protein is a preferred substrate of CDP II which is, as mentioned previously, also localized in the apical regions of the hyphae. These results indicate a putative functional relationship between these two proteins (spectrin-like protein and CDP II) in the dynamics of tip growth.

Keywords

Germ Tube Neurospora Crassa Immunogold Label Saprolegnia Calponin Homology Domain 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We gratefully acknowledge financial support from the Academic Society of Geneva (F. B.) and National Science Foundation Grant No. 3100-056786.99 (M. O.). Thanks are due to R. Strasser for interest in the project and encouragement, A. Cattaneo for technical assistance, and A. Fehr for secretarial assistance.

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • M. Cotado-Sampayo
    • 1
  • M. Ojha
    • 2
  • R. Ortega-Pérez
    • 1
  • M.-L. Chappuis
    • 1
  • F. Barja
    • 1
    Email author
  1. 1.Laboratory of Bioenergetics and MicrobiologyUniversity of GenevaJussy-GenevaSwitzerland
  2. 2.Department of BiochemistryUniversity of GenevaSwitzerland

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