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Immunolocalization of myosin in intact and wounded cells of the green alga Ernodesmis verticillata (Kützing) Borgesen

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Abstract

Myosin localization was examined in the coenocytic green alga E. verticillata using indirect immunofluorescence microscopy. A polyclonal antibody affinity-purified against the heavy chain of slime-mold myosin recognizes a 220000 to 230000 Mr protein that electrophoretically migrates slightly behind rabbit myosin. A second polypeptide of 85000 Mr is also consistently detected in immunoblots, indicating that two forms of myosin-like proteins may be present in these cells. In intact cells, myosin immunofluorescence is present on the chloroplast surfaces, in nuclei and in cytoplasmic strands between plastids. Myosin labeling also occurs in association with pyrenoids primarily in apical chloroplasts. During wound-induced cytoplasmic contractions, myosin is localized near the plasma membrane in longitudinal arrays superimposed over a reticulate pattern of fluorescence; both these patterns become apparent upon wounding. Double-label immunofluorescence of actin and myosin demonstrates that these arrays represent the longitudinal bundles of actin microfilaments and the actin-containing reticulum, the former being directly associated with contraction in these cells. These results indicate that both actin and myosin are associated with contractility in Ernodesmis, probably representing the apparatus and “molecular motor”, respectively, which effect motility.

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Abbreviations

DIC:

Nomarski differential interference contrast microscopy

FITC:

fluorescein isothiocyanate

MF(s):

microfllament(s)

TCA:

trichloroacetic acid

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Authors and Affiliations

  1. Department of Botany, University of Texas, 78713, Austin, TX, USA

    J. W. La Claire II

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The author is especially grateful to Professor J.A. West (University of California, Berkeley, USA) for the original algal isolates; Professor D.L. Herrin and Dr. R.H. Goddard (University of Texas) for use of the ultracentrifuge and for help with the TCA preparations, respectively; to Professor S.S. Brown (University of Michigan, Ann Arbor, USA) for help with the affinity purifications; and to Professor J.A. Spudich (Stanford University School of Medicine) and all the members of his laboratory for their warm hospitality, patient tutelage and generous support during my research leave. Special gratitude is extended to Dr. S. Ravid and K. Niebling (Stanford University School of Medicine) for valuable suggestions regarding the antibody work, and to J. Tan (Stanford University School of Medicine) for providing purified myosin. The research leave was funded in part by a University of Texas Faculty Research Award. Portions of this work were supported by National Science Foundation grant DCB 88-10084 and U.S. Department of Agriculture grant 87-CRCR-1-2545.

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La Claire, J.W. Immunolocalization of myosin in intact and wounded cells of the green alga Ernodesmis verticillata (Kützing) Borgesen. Planta 184, 209–217 (1991). https://doi.org/10.1007/BF00197949

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