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Characterization of Neurospora crassa α-Actinin

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Abstract

α-Actinin, an actin-binding protein of the spectrin superfamily, is present in most eukaryotes except plants. It is composed of three domains: N-terminal CH-domains, C-terminal calcium-binding domain (with EF-hand motifs), and a central rod domain. We have cloned and expressed Neurospora crassa α-actinin as GST and GFP fusion proteins for biochemical characterization and in vivo localization, respectively. The intracellular localization pattern of α-actinin suggests that this protein is intimately associated with actin filaments and plays an important role in the processes of germination, hyphal elongation, septum formation, and conidiation. These functions were confirmed by the experiments on the effect of α-actinin gene deletion in N. crassa.

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References

  1. Addis MF, Rappelli P, Delogu G, Carta F, Cappuccinelli P, Fiori PL (1998) Cloning and molecular characterization of a cDNA clone coding for Trichomonas vaginalis alpha-actinin and intracellular localization of the protein. Infect Immun 66:4924–4931

    PubMed  CAS  Google Scholar 

  2. Araujo-Bazan L, Penalva MA, Espeso EA (2008) Preferential localization of the endocytic internalization machinery to hyphal tips underlies polarization of the actin cytoskeleton in Aspergillus nidulans. Mol Microbiol 67:891–905

    Article  PubMed  CAS  Google Scholar 

  3. Barja F, Chappuis ML, Turian G (1993) Differential effects of anticytoskeletal compounds on the localization and chemical patterns of actin in germinating conidia of Neurospora crassa. FEMS Microbiol Lett 107:261–266

    Article  PubMed  CAS  Google Scholar 

  4. Berepiki A, Lichius A, Shoji JY, Tilsner J, Read ND (2010) F-actin dynamics in Neurospora crassa. Eukaryot Cell 9:547–557

    Article  PubMed  CAS  Google Scholar 

  5. Broderick MJ, Winder SJ (2005) Spectrin, α-actinin, and dystrophin. Adv Protein Chem 70:203–246

    Article  PubMed  CAS  Google Scholar 

  6. Colot HV, Park G, Turner GE, Ringelberg C, Crew CM, Litvinkova L, Weiss RL, Borkovich KA, Dunlap JC (2006) A high-throughput gene knockout procedure for Neurospora reveals functions for multiple transcription factors. Proc Natl Acad Sci 103:10352–10357

    Article  PubMed  CAS  Google Scholar 

  7. Cotado-Sampayo M, Ojha M, Ortega Perez R, Chappuis ML, Barja F (2006) Proteolytic cleavage of a spectrin-related protein by calcium-dependent protease in Neurospora crassa. Curr Microbiol 53:311–316

    Article  PubMed  CAS  Google Scholar 

  8. Cotado-Sampayo M, Okenve Ramos P, Ortega Perez R, Ojha M, Barja F (2008) Specificity of commercial anti-spectrin antibody in the study of fungal and Oomycete spectrin: cross-reaction with proteins other than spectrin. Fungal Genet Biol 45:1008–1015

    Article  PubMed  CAS  Google Scholar 

  9. Davis RH, De Serres FJ (1970) Genetic and microbial research techniques for Neurospora crassa. Methods Enzymol 17:79–143

    Article  Google Scholar 

  10. Delgado-Álvarez DL, Callejas-Negrete OA, Gómez N, Freitag M, Roberson RW, Smith LG, Mouriño-Pérez RR (2010) Visualization of F-actin localization and dynamics with live cell markers in Neurospora crassa. Fungal Genet Biol 47:573–586

    Article  PubMed  Google Scholar 

  11. Dominguez R (2004) Actin-binding proteins: a unifying hypothesis. Trends Biochem Sci 29:572–578

    Article  PubMed  CAS  Google Scholar 

  12. Freitag M, Hickey PC, Raju NB, Selker EU, Read ND (2004) GFP as a tool to analyze the organization, dynamics and function of nuclei and microtubules in Neurospora crassa. Fungal Genet Biol 41:897–910

    Article  PubMed  CAS  Google Scholar 

  13. Harris SD, Hamer L, Sharpless KE, Hamer JE (1997) The Aspergillus nidulans sepA gene encodes an FH1/2 protein involved in cytokinesis and the maintenance of cellular polarity. EMBO J 16:3474–3483

    Article  PubMed  CAS  Google Scholar 

  14. Heath IB, Gupta G, Bai S (2000) Plasma membrane-adjacent actin filaments, but not microtubules, are essential for both polarization and hyphal tip morphogenesis in Saprolegnia ferax and Neurospora crassa. Fungal Genet Biol 30:45–62

    Article  PubMed  CAS  Google Scholar 

  15. Lee SH, Dominguez R (2010) Regulation of actin cytoskeleton dynamics in cells. Mol Cells 29:311–325

    Article  PubMed  CAS  Google Scholar 

  16. Lek M, North KN (2010) Are biological sensors modulated by their structural scaffolds? The role of the structural muscle proteins alpha-actinin-2 and alpha-actinin-3 as modulators of biological sensors. FEBS Lett 584:2974–2980

    Article  PubMed  CAS  Google Scholar 

  17. McGoldrick CA, Gruver C, May CS (1995) myoA of Aspergillus nidulans encodes myosin I required for secretion and polarized growth. J Cell Biol 128:577–587

    Article  PubMed  CAS  Google Scholar 

  18. Otey CA, Carpen O (2004) Alpha-actinin revisited: a fresh look at an old player. Cell Motil Cytoskeleton 58:104–111

    Article  PubMed  CAS  Google Scholar 

  19. Rasmussen CG, Glass NL (2007) Localization of RHO-4 indicates differential regulation of conidial versus vegetative septation in the filamentous fungus Neurospora crassa. Eukaryot Cell 6:1097–1107

    Article  PubMed  CAS  Google Scholar 

  20. Riquelme M, Reynaga-Pena CG, Gierz G, Bartnicki-Garcia S (1998) What determines growth direction in fungal hyphae? Fungal Genet Biol 24:101–109

    Article  PubMed  CAS  Google Scholar 

  21. Rivero F, Furukawa R, Fechheimer M, Noegel AA (1999) Three actin cross-linking proteins, the 34 kDa actin-bundling protein, α-actinin and gelation factor (ABP-120), have both unique and redundant roles in the growth and development of Dictyostelium. J Cell Sci 112:2737–2751

    PubMed  CAS  Google Scholar 

  22. Sharpless KE, Harris SD (2002) Functional characterization and localization of the Aspergillus nidulans formin SEPA. Mol Biol Cell 13:469–479

    Article  PubMed  CAS  Google Scholar 

  23. Sjöblom B, Salmazo A, D**ović-Carugo K (2008) Alpha-actinin structure and regulation. Cell Mol Life Sci 65:2688–2701

    Article  PubMed  Google Scholar 

  24. Taheri-Talesh N, Horio T, Araujo-Bazan L, Dou X, Espeso EA, Peñalva MA, Osmani AS, Oakley BR (2008) The tip growth apparatus of Aspergillus nidulans. Mol Biol Cell 19:1439–1449

    Article  PubMed  CAS  Google Scholar 

  25. Upadhyay S, Shaw BD (2008) The role of actin, fimbrin and endocytosis in growth of hyphae in Aspergillus nidulans. Mol Microbiol 68:609–705

    Article  Google Scholar 

  26. Virag A, Griffiths AJ (2004) A mutation in the Neurospora crassa actin gene results in multiple defects in tip growth and branching. Fungal Genet Biol 41:213–225

    Article  PubMed  CAS  Google Scholar 

  27. Virag A, Harris SD (2006) Functional characterization of Aspergillus nidulans homologues of Saccharomyces cerevisiae Spa2 and Bud6. Eukaryot Cell 5:881–895

    Article  PubMed  CAS  Google Scholar 

  28. Virel A, Backman L (2004) Molecular evolution and structure of α-actinin. Mol Biol Evol 21:1024–1031

    Article  PubMed  CAS  Google Scholar 

  29. Virel A, Backman L (2006) Characterization of Entamoeba histolytica alpha-actinin. Mol Biochem Parasitol 145:11–17

    Article  PubMed  CAS  Google Scholar 

  30. Virel A, Backman L (2007) A comparative and phylogenetic analysis of the α-actinin rod domain. Mol Biol Evol 10:2254–2265

    Article  Google Scholar 

  31. Virel A, Addario B, Backman L (2007) Characterization of Entamoeba histolytica alpha-actinin 2. Mol Biochem Parasitol 154:82–89

    Article  PubMed  CAS  Google Scholar 

  32. Vogel HJ (1956) A convenient growth medium for Neurospora (medium N). Microbiol Genet Bull 13:42–43

    Google Scholar 

  33. Wang J, Hu H, Wang S, Shi J, Chen S, Wei H, Xu X, Lu L (2009) The important role of Actinin-like protein (AcnA) in cytokinesis and apical dominance of hyphal cells in Aspergillus nidulans. Microbiology 155:2714–2725

    Article  PubMed  CAS  Google Scholar 

  34. Wu JQ, Bahler J, Pringle JR (2001) Roles of a fimbrin and an α-actinin like protein in fission yeast cell polarization and cytokinesis. Mol Biol Cell 12:1061–1077

    PubMed  CAS  Google Scholar 

  35. Yamashita RA, Osherov N, May GS (2000) Localization of wild type and mutant class I myosin proteins in Aspergillus nidulans using GFP-fusion proteins. Cell Motil Cytoskeleton 45:163–172

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We gratefully acknowledge financial support from the Academic Society of Geneva (F. B.). Thanks are due to R. Strasser for interest in the project and encouragement, M.-L. Chappuis for technical assistance and A. Fehr for secretarial assistance.

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Authors and Affiliations

  1. Department of Botany and Plant Biology, University of Geneva, Chemin des Embrouchis 10, 1254, Jussy, Geneva, Switzerland

    Marta Cotado-Sampayo, Ruben Ortega Pérez & François Barja

  2. Department of Biochemistry, University of Geneva, Quai Ernest-Ansermet 30, 1211, Geneva 4, Switzerland

    Mukti Ojha & Carole Seum

Authors
  1. Marta Cotado-Sampayo
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  2. Ruben Ortega Pérez
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  3. Mukti Ojha
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  4. Carole Seum
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  5. François Barja
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Correspondence to François Barja.

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Cotado-Sampayo, M., Ortega Pérez, R., Ojha, M. et al. Characterization of Neurospora crassa α-Actinin. Curr Microbiol 63, 100–105 (2011). https://doi.org/10.1007/s00284-011-9954-9

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  • DOI: https://doi.org/10.1007/s00284-011-9954-9

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