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Mol Biol Cell. 2019 Oct 15;30(22):2827-2837. doi: 10.1091/mbc.E19-03-0141. Epub 2019 Sep 18.

The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins.

Author information

1
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160.
2
Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.
3
Department of Ophthalmology, University of Kansas Medical Center, Kansas City, KS 66160.

Abstract

The green alga Chlamydomonas reinhardtii is a leading model system to study photosynthesis, cilia, and the generation of biological products. The cytoskeleton plays important roles in all of these cellular processes, but to date, the filamentous actin network within Chlamydomonas has remained elusive. By optimizing labeling conditions, we can now visualize distinct linear actin filaments at the posterior of the nucleus in both live and fixed vegetative cells. Using in situ cryo-electron tomography, we confirmed this localization by directly imaging actin filaments within the native cellular environment. The fluorescently labeled structures are sensitive to the depolymerizing agent latrunculin B (Lat B), demonstrating the specificity of our optimized labeling method. Interestingly, Lat B treatment resulted in the formation of a transient ring-like filamentous actin structure around the nucleus. The assembly of this perinuclear ring is dependent upon a second actin isoform, NAP1, which is strongly up-regulated upon Lat B treatment and is insensitive to Lat B-induced depolymerization. Our study combines orthogonal strategies to provide the first detailed visual characterization of filamentous actins in Chlamydomonas, allowing insights into the coordinated functions of two actin isoforms expressed within the same cell.

PMID:
31532705
DOI:
10.1091/mbc.E19-03-0141

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