EFFECT OF NON-MUSCLE TROPOMYOSIN ISOFORMS ENCODED BY THE TPM1 GENE ON COFILIN-1 ACTIVITY TOWARD ACTIN FILAMENTS

Cover Page

Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription Access

Abstract

The actin cytoskeleton is an integral participant in a large number of cellular processes, such as organelle transport, motility, contractility, exocytosis and endocytosis. At the same time, it plays an important role in pathological processes, for example, malignant invasion of cancer cells. Actin-binding proteins, in particular proteins of the tropomyosin family (Tpm) and cofilins, participate in the remodeling of the actin cytoskeleton. For the study, we selected the least studied Tpm isoforms expressed from the TPM1 gene – Tpm1.7, Tpm1.8, and Tpm1.9 – as well as the more well-known Tpm1.1 and Tpm1.6. In this work, we studied the mutual influence of these Tpm isoforms and cofilin-1 (cof-1) on the dynamics of the actin cytoskeleton. Using the method of co-precipitation of these proteins, it was shown that Tpm1.7, Tpm1.8 and Tpm1.9 isoforms significantly interfere with cof-1 binding to the F-actin surface. Viscometry was used to evaluate the depolymerizing and severing effect of cof-1 on actin filament. Isoforms Tpm1.1, Tpm1.8, and Tpm1.6 effectively prevented the depolymerizing/severing effect of cof-1, while the protective effect of Tpm1.7 and Tpm1.9 was less pronounced. The rhodamine-phalloidin release assay was used to analyze conformational changes in F-actin induced by cof-1. All studied Tpm isoforms effectively prevented the effect of cof-1 on actin filament. The study showed that TPM1 gene products generally have an inhibitory effect on cof-1 activity in relation to the dynamics of actin filament polymerization/depolymerization. Such properties of Tpm isoforms may be important for the formation of specific intracellular populations of actin filaments.

About the authors

S. G Roman

Research Center of Biotechnology of the Russian Academy of Sciences

Moscow, Russia

A. V Slushchev

Research Center of Biotechnology of the Russian Academy of Sciences

Moscow, Russia

V. V Nefedova

Research Center of Biotechnology of the Russian Academy of Sciences

Moscow, Russia

A. M Matyushenko

Research Center of Biotechnology of the Russian Academy of Sciences

Email: ammatyushenko@mail.ru
Moscow, Russia

References

  1. Blanchoin, L., Boujemaa-Paterski, R., Sykes, C., and Plastino, J. (2014) Actin dynamics, architecture, and mechanics in cell motility, Physiol. Rev., 94, 235-263, https://doi.org/10.1152/physrev.00018.2013.
  2. Pollard, T. D., and Cooper, J. A. (2009) Actin, a central player in cell shape and movement, Science, 326, 1208-1212, https://doi.org/10.1126/science.1175862.
  3. Katsuta, H., Sokabe, M., and Hirata, H. (2024) From stress fiber to focal adhesion: a role of actin crosslinkers in force transmission, Front. Cell Dev. Biol., 12, 1444827, https://doi.org/10.3389/fcell.2024.1444827.
  4. Carlier, M.-F., and Shekhar, S. (2017) Global treadmilling coordinates actin turnover and controls the size of actin networks, Nat. Rev. Mol. Cell Biol., 18, 389-401, https://doi.org/10.1038/nrm.2016.172.
  5. Chaffer, C. L., San Juan, B. P., Lim, E., and Weinberg, R. A. (2016) EMT, cell plasticity and metastasis, Cancer Metastasis Rev., 35, 645-654, https://doi.org/10.1007/s10555-016-9648-7.
  6. Fife, C. M., McCarroll, J. A., and Kavallaris, M. (2014) Movers and shakers: cell cytoskeleton in cancer metastasis, Br. J. Pharmacol., 171, 5507-5523, https://doi.org/10.1111/bph.12704.
  7. Gibieza, P., and Petrikaitis, V. (2021) The regulation of actin dynamics during cell division and malignancy, Am. J. Cancer Res., 11, 4050-4069.
  8. Khaitlina, S. Y. (2014) Intracellular transport based on actin polymerization, Biochemistry (Moscow), 79, 917-927, https://doi.org/10.1134/S0006297914090089.
  9. Bamburg, J. R. (1999) Proteins of the ADF/cofilin family: essential regulators of actin dynamics, Annu. Rev. Cell Dev. Biol., 15, 185-230, https://doi.org/10.1146/annurev.cellbio.15.1.185.
  10. Carlier, M.-F., Laurent, V., Santolini, J., Melki, R., Didry, D., Xia, G.-X., Hong, Y., Chua, N.-H., and Pantaloni, D. (1997) Actin depolymerizing factor (ADF/cofilin) enhances the rate of filament turnover: implication in actin-based motility, J. Cell Biol., 136, 1307-1322, https://doi.org/10.1083/jcb.136.6.1307.
  11. Blanchoin, L., and Pollard, T. D. (1998) Interaction of actin monomers with acanthamoeba ectophorin (ADF/cofilin) and profilin, J. Biol. Chem., 273, 25106-25111, https://doi.org/10.1074/jbc.273.39.25106.
  12. De La Cruz, E. M. (2009) How cofilin severs an actin filament, Biophys. Rev., 1, 51-59, https://doi.org/10.1007/s12551-009-0008-5.
  13. McCullough, B. R., Grintsevich, E. E., Chen, C. K., Kang, H., Hutchison, A. L., Henn, A., Cao, W., Suarez, C., Martiel, J.-L., Blanchoin, L., Reisler, E., and De La Cruz, E. M. (2011) Cofilin-linked changes in actin filament flexibility promote severing, Biophys. J., 101, 151-159, https://doi.org/10.1016/j.bpj.2011.05.049.
  14. Galkin, V. E., Orlova, A., Kudryashov, D. S., Solodukhin, A., Reisler, E., Schröder, G. F., and Egelman, E. H. (2011) Remodeling of actin filaments by ADF/cofilin proteins, Proc. Natl. Acad. Sci. USA, 108, 20568-20572, https://doi.org/10.1073/pnas.1110109108.
  15. Prochniewicz, E., Janson, N., Thomas, D. D., and De La Cruz, E. M. (2005) Cofilin increases the torsional flexibility and dynamics of actin filaments, J. Mol. Biol., 353, 990-1000, https://doi.org/10.1016/j.jmb.2005.09.021.
  16. McGough, A., Pope, B., Chiu, W., and Weeds, A. (1997) Cofilin changes the twist of f-actin: implications for actin filament dynamics and cellular function, J. Cell Biol., 138, 771-781, https://doi.org/10.1083/jcb.138.4.771.
  17. McCullough, B. R., Blanchoin, L., Martiel, J.-L., and De La Cruz, E. M. (2008) Cofilin increases the bending flexibility of actin filaments: implications for severing and cell mechanics, J. Mol. Biol., 381, 550-558, https://doi.org/10.1016/j.jmb.2008.05.055.
  18. Dedova, I. V., Nikolaeva, O. P., Mikhailova, V. V., Dos Remedios, C. G., and Levitsky, D. I. (2004) Two opposite effects of cofilin on the thermal unfolding of F-actin: a differential scanning calorimetric study, Biophys. Chem., 110, 119-128, https://doi.org/10.1016/j.bpc.2004.01.009.
  19. Pavlov, D., Muhlrad, A., Cooper, J., Wear, M., and Reisler, E. (2007) Actin filament severing by cofilin, J. Mol. Biol., 365, 1350-1358, https://doi.org/10.1016/j.jmb.2006.10.102.
  20. Bamburg, J. R., and Bernstein, B. W. (2016) Actin dynamics and cofilin-actin rods in Alzheimer disease, Cytoskeleton, 73, 477-497, https://doi.org/10.1002/cm.21282.
  21. Kanellos, G., and Frame, M. C. (2016) Cellular functions of the ADF/cofilin family at a glance, J. Cell Sci., 129, 3211-3218, https://doi.org/10.1242/jcs.187849.
  22. Ohashi, K. (2015) Roles of cofilin in development and its mechanisms of regulation, Dev. Growth Differ., 57, 275-290, https://doi.org/10.1111/dgd.12213.
  23. Thirion, C., Stucka, R., Mendel, B., Gruhler, A., Jaksch, M., Nowak, K. J., Binz, N., Laing, N. G., and Lochmüller, H. (2001) Characterization of human muscle type cofilin (CFL2) in normal and regenerating muscle, Eur. J. Biochem., 268, 3473-3482, https://doi.org/10.1046/j.1432-1327.2001.02247.x.
  24. Kremneva, E., Makkonen, M. H., Skwarek-Maruszewska, A., Gateva, G., Michelot, A., Dominguez, R., and Lappalainen, P. (2014) Cofilin-2 controls actin filament length in muscle sarcomeres, Dev. Cell, 31, 215-226, https://doi.org/10.1016/j.devcel.2014.09.002.
  25. Sexton, J. A., Potchernikov, T., Bibeau, J. P., Casanova-Sepúlveda, G., Cao, W., Lou, H. J., Boggon, T. J., De La Cruz, E. M., and Turk, B. E. (2024) Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail, Nat. Commun., 15, 1426, https://doi.org/10.1038/s41467-024-45878-9.
  26. Moriyama, K., Iida, K., and Yahara, I. (1996) Phosphorylation of Ser-3 of cofilin regulates its essential function on actin, Genes Cells, 1, 73-86, https://doi.org/10.1046/j.1365-2443.1996.05005.x.
  27. Niwa, R., Nagata-Ohashi, K., Takeichi, M., Mizuno, K., and Uemura, T. (2002) Control of actin reorganization by Slingshot, a family of phosphatases that dephosphorylate ADF/cofilin, Cell, 108, 233-246, https://doi.org/10.1016/S0092-8674(01)00638-9.
  28. Ono, S. (2018) Functions of actin-interacting protein 1 (AIP1)/WD repeat protein 1 (WDR1) in actin filament dynamics and cytoskeletal regulation, Biochem. Biophys. Res. Commun., 506, 315-322, https://doi.org/10.1016/j.bbrc.2017.10.096.
  29. Ostrowska-Podhorodecka, Z., Śliwińska, M., Reisler, E., and Moraczewska, J. (2020) Tropomyosin isoforms regulate cofilin 1 activity by modulating actin filament conformation, Arch. Biochem. Biophys., 682, 108280, https://doi.org/10.1016/j.abb.2020.108280.
  30. Geeves, M. A., Hitchcock-Defregori, S. E., and Gunning, P. W. (2015) A systematic nomenclature for mammalian tropomyosin isoforms, J. Muscle Res. Cell Motil., 36, 147-153, https://doi.org/10.1007/s10974-014-9389-6.
  31. Nevzorov, I. A., and Levitsky, D. I. (2011) Tropomyosin: double helix from the protein world, Biochemistry (Moscow), 76, 1507-1527, https://doi.org/10.1134/S0006297911130098.
  32. Janco, M., Bonello, T. T., Byun, A., Coster, A. C. F., Lebhar, H., Dedova, I., Gunning, P. W., and Böcking, T. (2016) The impact of tropomyosins on actin filament assembly is isoform specific, BioArchitecture, 6, 61-75, https://doi.org/10.1080/19490992.2016.1201619.
  33. Gray, K. T., Kostyukova, A. S., and Fath, T. (2017) Actin regulation by tropomodulin and tropomyosin in neuronal morphogenesis and function, Mol. Cell. Neurosci., 84, 48-57, https://doi.org/10.1016/j.mcn.2017.04.002.
  34. Gunning, P. W., Hardeman, E. C., Lappalainen, P., and Mulvihill, D. P. (2015) Tropomyosin – master regulator of actin filament function in the cytoskeleton, J. Cell Sci., 128, 2965-2974, https://doi.org/10.1242/jcs.172502.
  35. Sung, L. A., Gao, K. M., Yee, L. J., Temm-Grove, C. J., Helfman, D. M., Lin, J. J., and Mehrpouryan, M. (2000) Tropomyosin isoform 5b is expressed in human erythrocytes: implications of tropomodulin-TM5 or tropomodulin-TM5 complexes in the protofilament and hexagonal organization of membrane skeletons, Blood, 95, 1473-1480, https://doi.org/10.1182/blood.v95.4.1473.004k50_1473_1480.
  36. Gokhin, D. S., and Fowler, V. M. (2016) Feisty filaments: actin dynamics in the red blood cell membrane skeleton, Curr. Opin. Hematol., 23, 206-214, https://doi.org/10.1097/MOH.0000000000000227.
  37. Fowler, V. M. (2013) The human erythrocyte plasma membrane, Curr. Top. Membr., 72, 39-88, https://doi.org/10.1016/B978-0-12-417027-8.00002-7.
  38. Sui, Z., Gokhin, D. S., Nowak, R. B., Guo, X., An, X., and Fowler, V. M. (2017) Stabilization of F-actin by tropomyosin isoforms regulates the morphology and mechanical behavior of red blood cells, Mol. Biol. Cell, 28, 2531-2542, https://doi.org/10.1091/mbc.e16-10-0699.
  39. Hughes, J. A. I., Cooke-Yarborough, C. M., Chadwick, N. C., Schevzov, G., Arbuckle, S. M., Gunning, P., and Weinberger, R. P. (2003) High-molecular-weight tropomyosins localize to the contractile rings of dividing CNS cells but are absent from malignant pediatric and adult CNS tumors, Glia, 42, 25-35, https://doi.org/10.1002/glia.10174.
  40. Dalby-Payne, J. R., O’Loughlin, E. V., and Gunning, P. (2003) Polarization of specific tropomyosin isoforms in gastrointestinal epithelial cells and their impact on CFTR at the apical surface, Mol. Biol. Cell, 14, 4365-4375, https://doi.org/10.1091/mbc.e03-03-0169.
  41. Schevzov, G., Whittaker, S. P., Fath, T., Lin, J. J., and Gunning, P. W. (2011) Tropomyosin isoforms and reagents, Bioarchitecture, 1, 135-164, https://doi.org/10.4161/bioa.1.4.17897.
  42. Lin, J. J.-C., Eppinga, R. D., Warren, K. S., and McCrae, K. R. (2008) Human tropomyosin isoforms in the regulation of cytoskeleton functions, Adv. Exp. Med. Biol., 644, 201-222, https://doi.org/10.1007/978-0-387-85766-4_16.
  43. Manstein, D. J., and Mulvihill, D. P. (2016) Tropomyosin-mediated regulation of cytoplasmic myosins, Traffic, 17, 872-877, https://doi.org/10.1111/tra.12399.
  44. Blanchoin, L., Pollard, T. D., and Hitchcock-Defregori, S. E. (2001) Inhibition of the Arp2/3 complex-nucleated actin polymerization and branch formation by tropomyosin, Curr. Biol., 11, 1300-1304, https://doi.org/10.1016/S0960-9822(01)00395-5.
  45. Kuhn, T. B., and Bamburg, J. R. (2008) Tropomyosin and ADF/cofilin as collaborators and competitors, Adv. Exp. Med. Biol., 644, 232-249, https://doi.org/10.1007/978-0-387-85766-4_18.
  46. Robaszkiewicz, K., Ostrowska, Z., Marchiewicz, K., and Moraczewska, J. (2016) Tropomyosin isoforms differentially modulate the regulation of actin filament polymerization and depolymerization by cofilins, FEBS J., 283, 723-737, https://doi.org/10.1111/febs.13626.
  47. Robaszkiewicz, K., Śliwińska, M., and Moraczewska, J. (2020) Regulation of actin filament length by muscle isoforms of tropomyosin and cofilin, Int. J. Mol. Sci., 21, 4285, https://doi.org/10.3390/ijms21124285.
  48. Robaszkiewicz, K., Wróbel, J., and Moraczewska, J. (2023) Troponin and a myopathy-linked mutation in TPM3 inhibit cofilin-2-induced thin filament depolymerization, Int. J. Mol. Sci., 24, 16457, https://doi.org/10.3390/ijms242216457.
  49. Ostrowska, Z., Robaszkiewicz, K., and Moraczewska, J. (2017) Regulation of actin filament turnover by cofilin-1 and cytoplasmic tropomyosin isoforms, Biochim. Biophys. Acta Prot. Proteomics, 1865, 88-98, https://doi.org/10.1016/j.bbapap.2016.09.019.
  50. Selvaraj, M., Kokate, S. B., Reggiano, G., Kogan, K., Kotila, T., Kremneva, E., DiMaio, F., Lappalainen, P., and Huiskonen, J. T. (2023) Structural basis underlying specific biochemical activities of non-muscle tropomyosin isoforms, Cell Rep., 42, 111900, https://doi.org/10.1016/j.celrep.2022.111900.
  51. Gateva, G., Kremneva, E., Reindl, T., Kotila, T., Kogan, K., Gressin, L., Gunning, P. W., Manstein, D. J., Michelot, A., and Lappalainen, P. (2017) Tropomyosin isoforms specify functionally distinct actin filament populations in vitro, Curr. Biol., 27, 705-713, https://doi.org/10.1016/j.cub.2017.01.018.
  52. Marchenko, M., Nefedova, V., Artemova, N., Kleymenov, S., Levitsky, D., and Matyushenko, A. (2021) Structural and functional peculiarities of cytoplasmic tropomyosin isoforms, the products of TPM1 and TPM4 genes, Int. J. Mol. Sci., 22, 5141, https://doi.org/10.3390/ijms22105141.
  53. Lapshina, K. K., Nefedova, V. V., Nabiev, S. R., Roman, S. G., Shchepkin, D. V., Kopylova, G. V., Kochurova, A. M., Beldija, E. A., Kleymenov, S. Y., Levitsky, D. I., and Matyushenko, A. M. (2024) Functional and structural properties of cytoplasmic tropomyosin isoforms Tpm1.8 and Tpm1.9, Int. J. Mol. Sci., 25, 6873, https://doi.org/10.3390/ijms25136873.
  54. Monteiro, P. B., Lataro, R. C., Ferro, J. A., and Reinach, F. de C. (1994) Functional alpha-tropomyosin produced in Escherichia coli. A dipeptide extension can substitute the amino-terminal acetyl group, J. Biol. Chem., 269, 10461-10466, https://doi.org/10.1016/S0021-9258(17)34082-6.
  55. Umeki, N., Hirose, K., and Uyeda, T. Q. P. (2016) Cofilin-induced cooperative conformational changes of actin subunits revealed using cofilin-actin fusion protein, Sci. Rep., 6, 20406, https://doi.org/10.1038/srep20406.
  56. Pardee, J. D., and Aspudich, J. (1982) Purification of muscle actin, Methods Enzymol., 85, 164-181, https://doi.org/10.1016/0076-6879(82)85020-9.
  57. Bobkov, A. A., Muhlrad, A., Shvetsov, A., Benchatar, S., Scoville, D., Almo, S. C., and Reisler, E. (2004) Cofilin (ADF) affects lateral contacts in F-actin, J. Mol. Biol., 337, 93-104, https://doi.org/10.1016/j.jmb.2004.01.014.
  58. Chou, S. Z., and Pollard, T. D. (2020) Cryo-electron microscopy structures of pyrene-labeled ADP-Pi- and ADP-actin filaments, Nat. Commun., 11, 5897, https://doi.org/10.1038/s41467-020-19762-1.
  59. Muhlrad, A., Ringel, I., Pavlov, D., Peyser, Y. M., and Reisler, E. (2006) Antagonistic effects of cofilin, beryllium fluoride complex, and phalloidin on subdomain 2 and nucleotide-binding cleft in F-actin, Biophys. J., 91, 4490-4499, https://doi.org/10.1529/biophysj.106.087767.
  60. Gunning, P., O'Neill, G., and Hardeman, E. (2008) Tropomyosin-based regulation of the actin cytoskeleton in time and space, Physiol. Rev., 88, 1-35, https://doi.org/10.1152/physrev.00001.2007.
  61. Kis-Bieskei, N., Vig, A., Nyirrai, M., Bugyi, B., and Talian, G. C. (2013) Purification of tropomyosin Br-3 and SNM1 and characterization of their interactions with actin, Cytoskeleton, 70, 755-765, https://doi.org/10.1002/cm.21143.
  62. Moraczewska, J., Nicholson-Flynn, K., and Hitchcock-DeGregori, S. E. (1999) The ends of tropomyosin are major determinants of actin affinity and myosin subfragment 1-induced binding to F-actin in the open state, Biochemistry, 38, 15885-15892, https://doi.org/10.1021/bi991816j.
  63. Matyushenko, A. M., Koubassova, N. A., Shchepkin, D. V., Kopylova, G. V., Nabiev, S. R., Nikitina, L. V., Bershitsky, S. Y., Levitsky, D. I., and Tsaturyan, A. K. (2019) The effects of cardiomyopathy-associated mutations in the head-to-tail overlap junction of α-tropomyosin on its properties and interaction with actin, Int. J. Biol. Macromol., 125, 1266-1274, https://doi.org/10.1016/j.ijbiomac.2018.09.105.
  64. Matyushenko, A. M., Shchepkin, D. V., Kopylova, G. V., Bershitsky, S. Y., and Levitsky, D. I. (2020) Unique functional properties of slow skeletal muscle tropomyosin, Biochimie, 174, 1-8, https://doi.org/10.1016/j.biochi.2020.03.013.

Supplementary files

Supplementary Files
Action
1. JATS XML
Download

Copyright (c) 2025 Russian Academy of Sciences

Согласие на обработку персональных данных с помощью сервиса «Яндекс.Метрика»

1. Я (далее – «Пользователь» или «Субъект персональных данных»), осуществляя использование сайта https://journals.rcsi.science/ (далее – «Сайт»), подтверждая свою полную дееспособность даю согласие на обработку персональных данных с использованием средств автоматизации Оператору - федеральному государственному бюджетному учреждению «Российский центр научной информации» (РЦНИ), далее – «Оператор», расположенному по адресу: 119991, г. Москва, Ленинский просп., д.32А, со следующими условиями.

2. Категории обрабатываемых данных: файлы «cookies» (куки-файлы). Файлы «cookie» – это небольшой текстовый файл, который веб-сервер может хранить в браузере Пользователя. Данные файлы веб-сервер загружает на устройство Пользователя при посещении им Сайта. При каждом следующем посещении Пользователем Сайта «cookie» файлы отправляются на Сайт Оператора. Данные файлы позволяют Сайту распознавать устройство Пользователя. Содержимое такого файла может как относиться, так и не относиться к персональным данным, в зависимости от того, содержит ли такой файл персональные данные или содержит обезличенные технические данные.

3. Цель обработки персональных данных: анализ пользовательской активности с помощью сервиса «Яндекс.Метрика».

4. Категории субъектов персональных данных: все Пользователи Сайта, которые дали согласие на обработку файлов «cookie».

5. Способы обработки: сбор, запись, систематизация, накопление, хранение, уточнение (обновление, изменение), извлечение, использование, передача (доступ, предоставление), блокирование, удаление, уничтожение персональных данных.

6. Срок обработки и хранения: до получения от Субъекта персональных данных требования о прекращении обработки/отзыва согласия.

7. Способ отзыва: заявление об отзыве в письменном виде путём его направления на адрес электронной почты Оператора: info@rcsi.science или путем письменного обращения по юридическому адресу: 119991, г. Москва, Ленинский просп., д.32А

8. Субъект персональных данных вправе запретить своему оборудованию прием этих данных или ограничить прием этих данных. При отказе от получения таких данных или при ограничении приема данных некоторые функции Сайта могут работать некорректно. Субъект персональных данных обязуется сам настроить свое оборудование таким способом, чтобы оно обеспечивало адекватный его желаниям режим работы и уровень защиты данных файлов «cookie», Оператор не предоставляет технологических и правовых консультаций на темы подобного характера.

9. Порядок уничтожения персональных данных при достижении цели их обработки или при наступлении иных законных оснований определяется Оператором в соответствии с законодательством Российской Федерации.

10. Я согласен/согласна квалифицировать в качестве своей простой электронной подписи под настоящим Согласием и под Политикой обработки персональных данных выполнение мною следующего действия на сайте: https://journals.rcsi.science/ нажатие мною на интерфейсе с текстом: «Сайт использует сервис «Яндекс.Метрика» (который использует файлы «cookie») на элемент с текстом «Принять и продолжить».