期刊
JOURNAL OF CELL SCIENCE
卷 135, 期 18, 页码 -出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.260288
关键词
Actin; Live imaging; Tropomyosin; mNeonGreen; Nanobody; Cytokinesis
类别
资金
- Wellcome Trust [203276/Z/16/Z, WT101885MA, 220790/Z/20/Z]
- Schweizerischer Nationalfonds zur Forderung der Wissenschaftlichen Forschung (Swiss National Science Foundation) [310030B_176396]
- European Research Council
- Biotechnology and Biological Sciences Research Council [BB/S003789/1, BB/T000481/1]
- Wellcome Trust DBT India Alliance [IA/I/21/1/505633]
- DST Science and Engineering Research Board (SERB) [SRG/2021/001600]
- GATE fellowship
- University of Warwick
- Wellcome Trust [220790/Z/20/Z] Funding Source: Wellcome Trust
In this study, a new tool for observing the dynamics of tropomyosin was reported and successfully applied in different organisms. The study revealed the important role of tropomyosin in cytoskeletal regulation and its functions in cell division and mating processes.
Tropomyosins are structurally conserved a-helical coiled-coil proteins that bind along the length of filamentous actin (F-actin) in fungi and animals. Tropomyosins play essential roles in the stability of actin filaments and in regulating myosin II contractility. Despite the crucial role of tropomyosin in actin cytoskeletal regulation, in vivo investigations of tropomyosin are limited, mainly due to the suboptimal live-cell imaging tools currently available. Here, we report on an mNeonGreen (mNG)-tagged tropomyosin, with native promoter and linker length configuration, that clearly reports tropomyosin dynamics in Schizosaccharomyces pombe (Cdc8), Schizosaccharomyces japonicus (Cdc8) and Saccharomyces cerevisiae (Tpm1 and Tpm2). We also describe a fluorescent probe to visualize mammalian tropomyosin (TPM2 isoform). Finally, we generated a camelid nanobody against S. pombe Cdc8, which mimics the localization of mNG-Cdc8 in vivo. Using these tools, we report the presence of tropomyosin in previously unappreciated patch-like structures in fission and budding yeasts, show flow of tropomyosin (F-actin) cables to the cytokinetic actomyosin ring and identify rearrangements of the actin cytoskeleton during mating. These powerful tools and strategies will aid better analyses of tropomyosin and F-actin cables in vivo.
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