Parallel assembly of actin and tropomyosin, but not myosin II, during de novo actin filament formation in live mice

Journal of Cell Science
Andrius MasedunskasEdna C Hardeman

Abstract

Many actin filaments in animal cells are co-polymers of actin and tropomyosin. In many cases, non-muscle myosin II associates with these co-polymers to establish a contractile network. However, the temporal relationship of these three proteins in the de novo assembly of actin filaments is not known. Intravital subcellular microscopy of secretory granule exocytosis allows the visualisation and quantification of the formation of an actin scaffold in real time, with the added advantage that it occurs in a living mammal under physiological conditions. We used this model system to investigate the de novo assembly of actin, tropomyosin Tpm3.1 (a short isoform of TPM3) and myosin IIA (the form of non-muscle myosin II with its heavy chain encoded by Myh9) on secretory granules in mouse salivary glands. Blocking actin polymerization with cytochalasin D revealed that Tpm3.1 assembly is dependent on actin assembly. We used time-lapse imaging to determine the timing of the appearance of the actin filament reporter LifeAct-RFP and of Tpm3.1-mNeonGreen on secretory granules in LifeAct-RFP transgenic, Tpm3.1-mNeonGreen and myosin IIA-GFP (GFP-tagged MYH9) knock-in mice. Our findings are consistent with the addition of tropomyosin to actin fil...Continue Reading

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Citations

Aug 7, 2018·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Seham EbrahimRoberto Weigert
Jun 14, 2018·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Wen Yu WongKirsten H Limesand
May 6, 2019·Journal of Muscle Research and Cell Motility·Dietmar J MansteinPeter W Gunning
Jul 25, 2019·Journal of Cell Science·Joyce C M MeiringPeter W Gunning
Jul 9, 2020·Biophysics Reviews·Miro JancoPeter W Gunning
Jun 18, 2020·Cells·Pika Miklavc, Manfred Frick
Dec 7, 2020·Molecular Therapy : the Journal of the American Society of Gene Therapy·Nan WuBurton B Yang
Oct 22, 2019·Seminars in Cell & Developmental Biology·Edna C HardemanPeter W Gunning

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