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Identification and localization of calcium-dependent protease II inNeurospora crassa andUromyces appendiculatus

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Summary

The existence of Ca2+-dependent protease II in crude extracts ofNeurospora crassa andUromyces appendiculatus was demonstrated by immunoblotting using specific antibodies. In both extracts two immunoreacting bands were observed. The molecular mass of the major band inN. crassa corresponded to 37 kDa, while that inU. appendiculatus was 43 kDa, similar to that previously reported forAllomyces arbuscula. Immunofluorescence of the enzyme was predominantly localized in the apical regions of germlings and growing hyphae, suggesting a functional role for the enzyme in hyphal growth.

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References

  • Andersson M, Sjöstrand J, Karlsson J-O (1996) Calpains in the human lens: relations to membranes and possible role in cataract formation. Ophthalmic Res 28 (Suppl 1): 51–54

    Google Scholar 

  • Barja F, Nguyen Thi B-N, Turian G (1991a) Localization of actin and characterization of its isoforms in the hyphae ofNeurospora crassa. FEMS Microbiol Lett 77: 19–24

    Google Scholar 

  • —, Rossier C, Turian G (1991b) Actin microfilaments requirement in contrast with microtubules dispensability for germ tube outgrowth from conidia ofNeurospora crassa. Plant Physiol 10: 63–65

    Google Scholar 

  • —, Chappuis M-L, Turian G (1993) Differential effects of anticytoskeletal compounds on the localization and chemical patterns of actin in germinating conidia ofNeurospora crassa. FEMS Microbiol Lett 107: 261–266

    Google Scholar 

  • —, Corrêa A Jr, Staples RC, Hoch HC (1998) Microinjected antisenseInf24 oligonucleotides inhibit appressorium development inUromyces. Mycol Res 102: 1513–1518

    Google Scholar 

  • Billger M, Wallin M, Karlsson J-O (1988) Proteolysis of tubulin and microtubule-associated proteins 1 and 2 by calpain I and II: difference in sensitivity of assembled and disassembled microtubules. Cell Calcium 9: 33–44

    Google Scholar 

  • Burnette WM (1981) Western blotting electrophoresis transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and ratio-iodinated protein A. Anal Biochem 112: 195–203

    Google Scholar 

  • Corrêa A Jr, Hoch HC (1993) Microinjection of uredospore germlings ofUromyces appendiculatus. Exp Mycol 17: 241–252

    Google Scholar 

  • Croall DE, DeMartino GN (1991) Calcium-activated neutral protease (calpain) system: structure, function, and regulation. Physiol Rev 71: 813–847

    Google Scholar 

  • Denison SH, Orejas M, Arst HN Jr (1995) Signaling of ambient pH inAspergillus involves a cysteine protease. J Biol Chem 270: 26519–28522

    Google Scholar 

  • Fox JEB, Goll DE, Raynolds CC, Philips DR (1985) Identification of two proteins (actin-binding protein and P235) that are hydrolyzed by endogenous Ca2+-dependent protease during platelet aggregation. J Biol Chem 260: 1060–1066

    Google Scholar 

  • French RC, Graham CL, Gale AW, Long RK (1977) Structural and exposure time requirements for chemical stimulation of germination of uredospores ofUromyces phaseoli. J Agric Food Chem 25: 84–88

    Google Scholar 

  • Goll DE, Dayton WR, Singh J, Robson R (1991) Studies of the α-actinin/actin interaction in z disk by using calpain. J Biol Chem 266: 8501–8510

    Google Scholar 

  • Hoch HC, Tucker BE, Staples RC (1987) An intact microtubule cytoskeleton is necessary for mediation of the signal for cell differentiation inUromyces. Eur J Cell Biol 45: 209–218

    Google Scholar 

  • Huber D (1996) Protease calcium-dépendante (CANP II) chezAllomyces et possibles interactions avec le cytosquelette. Thèse no 2854, Université de Genève, Geneva, Switzerland

    Google Scholar 

  • —, Ojha M (1994) Immunocytochemical localization of Ca2+-dependent protease inAllomyces arbuscula. FEBS Lett 341: 268–272

    Google Scholar 

  • —, Ojha M, Turian G (1994) In vivo and in situ proteolysis of some cytoskeletal proteins by the Ca2+-dependent protease inAllomyces. Experientia 50: S23 (A88)

    Google Scholar 

  • Hyde GJ, Heath IB (1997) Ca2+ gradients in hyphae and branches ofSaprolegnia ferax. Fungal Genet Biol 21: 238–251

    Google Scholar 

  • Johnson GVW, Guttmann RP (1997) Calpains: intact and active? BioEssays 19: 1011–1018

    Google Scholar 

  • Kwon YH, Hoch HC, Staples RC (1991) Cytoskeletal organization inUromyces urediospore germling apices during appressorium formation. Protoplasma 165: 37–50

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  • Molinari M, Anagli J, Carafoli E (1994) Ca2+-activated neutral protease is active in the erythrocyte membrane in its nonautolyzed 80-kDa form. J Biol Chem 269: 27992–27995

    Google Scholar 

  • Moloshok TD, Leinhos GME, Staples RC, Hoch HC (1993) The autogenic extracellular environment ofUromyces appendiculatus urediospore germlings. Mycologia 85: 392–400

    Google Scholar 

  • Mykles DL, Skinner DM (1986) Four Ca2+-dependent proteinase activities isolated from crustacean muscle differ in size, net charge, and sensitivity to Ca2+and inhibitors. J Biol Chem 261: 9865–9871

    Google Scholar 

  • Ojha M (1990) A novel Ca2+-activated protease from an aquatic fungus,Allomyces arbuscula. In: O'Day DH (ed) Calcium as an intracellular messenger in eukaryotic microbes. American Society for Microbiology, Washington, DC, pp 192–211

    Google Scholar 

  • — (1996a) Calcium-dependent protease I fromAllomyces arbuscula. Biochem Biophys Res Commun 218: 22–29

    Google Scholar 

  • — (1996b) Purification, properties and developmental regulation of a Ca2+-independent serine-cysteine protease fromAllomyces arbuscula. Int J Biochem Cell Biol 28: 345–352

    Google Scholar 

  • —, Favre B (1991) In vitro and in vivo phosphorylation of calpain-like protease fromAllomyces arbuscula. Plant Sci 74: 35–44

    Google Scholar 

  • —, Wallace CJA (1988) Novel calcium-activated neutral protease from an aquatic fungus,Allomyces arbuscula. J Bacteriol 170: 1254–1260

    Google Scholar 

  • —, Cattaneo A, Schwendimann B (1999) Comparative study of Ca2+-dependent proteases (CDP I and CDP II) fromAllomyces arbuscula. Biochimie 81: 765–770

    Google Scholar 

  • Pintér M, Friedrich P (1988) The calcium-dependent proteolytic system calpain-calpastatin inDrosophila melanogaster. Biochem J 253: 467–473

    Google Scholar 

  • Pontremoli S, Melloni E, Salamino F, Sparatore B, Viotti P, Michetti M, Horecker BL (1984) Cytosolic Ca2+-dependent neutral proteinases from rabbit liver: activation of the proenzyme by Ca2+ and substrate. Proc Natl Acad Sci USA 81: 53–56

    Google Scholar 

  • Safadi F, Mykles DL, Reddy ASN (1997) Partial purification and characterization for a Ca2+-dependent proteinase fromArabidopsis roots. Arch Biochem Biophys 348: 143–151

    Google Scholar 

  • Sorimachi H, Kimura S, Kinbara K, Kazama J, Takahashi M, Yajima H, Ishiura S, Sasagawa N, Nonaka I, Sugita H, Maruyama K, Suzuki K (1996) Structure and physiological functions of ubiquitous and tissue-specific calpain species. Adv Biophys 33: 101–122

    Google Scholar 

  • Spector T (1978) Refinement of the Coomassie blue method of protein quantitation. Anal Biochem 86: 142–146

    Google Scholar 

  • Springer JE, Azbill RD, Kennedy SE, George J, Geddes JW (1997) Rapid calpain I activation and cytoskeletal protein degradation following traumatic spinal cord injury: attenuation with riluzole pretreatment. J Neurochem 69: 1592–1600

    Google Scholar 

  • Turian G, Schönenberger I, Nguyen Thi BN (1992) Cytoskeletal involvement in the bipolar disjunction ofAllomyces gametangia. Biomed Lett 47: 215–219

    Google Scholar 

  • Vogel HJ (1956) A convenient growth medium forNeurospora (medium N). Microb Genet Bull 13: 42–43

    Google Scholar 

  • Zimmerman UJ, Schlaepfer WW (1984) Calcium-activated neutral protease (CANP) in brain and other tissues. Prog Neurobiol 23: 63–78

    Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Bioénergétique et de Microbiologie, Université de Genève, Geneva

    Francisco Barja, Y. Jaquet, R. Ortega Perez & M. Ojha

  2. Department of Plant Pathology, Cornell University, New York State Agricultural Experiment Station, Geneva, New York

    H. C. Hoch

Authors
  1. Francisco Barja
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  2. Y. Jaquet
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  3. R. Ortega Perez
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  4. H. C. Hoch
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  5. M. Ojha
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Correspondence to Francisco Barja.

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Barja, F., Jaquet, Y., Perez, R.O. et al. Identification and localization of calcium-dependent protease II inNeurospora crassa andUromyces appendiculatus . Protoplasma 210, 85–91 (1999). https://doi.org/10.1007/BF01314958

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  • DOI: https://doi.org/10.1007/BF01314958

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