Differential response of myofibrillar and cytoskeletal proteins in cells treated with phorbol myristate acetate

Z. Lin, James Eshleman, C. Grund, D. A. Fischman, T. Masaki, W. W. Franke, H. Holtzer

Research output: Contribution to journalArticle

Abstract

Muscle-specific and nonmuscle contractile protein isoforms responded in opposite ways to 12-o-tetradecanoyl phorbol-13-acetate (TPA). Loss of Z band density was observed in day-4-5 cultured chick myotubes after 2 h in the phorbol ester, TPA. By 5-10 h, most I-Z-I complexes were selectively deleted from the myofibril, although the A bands remained intact and longitudinally aligned. The deletion of I-Z-I complexes was inversely related to the appearance of numerous cortical, α-actinin containing bodies (CABs), transitory structures ~3.0 μm in diameter. Each CAB consisted of a filamentous core that costained with antibodies to α-actin and sarcomeric α-actinin. In turn each CAB was encaged by a discontinuous rim that costained with antibodies to vinculin and talin. Vimentin and desmin intermediate filaments and most cell organelles were excluded from the membrane-free CABs. These curious bodies disappeared over the next 10 h so that in 30-h myosacs all α-actin and sarcomeric α-actinin structures had been eliminated. On the other hand vinculin and talin adhesion plaques remained prominent even in 72-h myosacs. Disruption of the A bands was first initiated after 15-20 h in TPA (e.g., 15-20-h myosacs). Thick filaments of apparently normal length and structure were progressively released from A segments, and by 40 h all A bands had been broken down into enormous numbers of randomly dispersed, but still intact single thick filaments. This breakdown correlated with the formation of amorphous cytoplasmic aggregates which invariably colocalized antibodies to myosin heavy chain, MLC 1-3, myomesin, and C protein. Complete elimination of all immunoreactive thick filament proteins required 60-72 h of TPA exposure. The elimination of the thick filament-associated proteins did not involve the participation of vinculin or talin. In contrast to its effects on myofibrils, TPA did not induce the disassembly of the contractile proteins in stress fibers and microfilaments either in myosacs or in fibroblastic cells. Similarly, TPA, which rapidly induces the translocation of vinculin and talin to ectopic sites in many types of immortalized cells, had no gross effect on the adhesion plaques of myosacs, primary fibroblastic cells, or presumptive myoblasts. Clearly, the response to TPA of contractile protein and some cytoskeletal isoforms not only varies among phenotypes, but even within the domains of a given myotube the myofibrils respond one way, the stress fibers/microfilaments another.

Original languageEnglish (US)
Pages (from-to)1079-1091
Number of pages13
JournalJournal of Cell Biology
Volume108
Issue number3
StatePublished - 1989
Externally publishedYes

Fingerprint

Cytoskeletal Proteins
Tetradecanoylphorbol Acetate
Talin
Acetates
Vinculin
Actinin
Contractile Proteins
Myofibrils
Stress Fibers
Focal Adhesions
Skeletal Muscle Fibers
Actin Cytoskeleton
Antibodies
Actins
Protein Isoforms
Connectin
Desmin
Intermediate Filaments
Myosin Heavy Chains
Myoblasts

ASJC Scopus subject areas

  • Cell Biology

Cite this

Lin, Z., Eshleman, J., Grund, C., Fischman, D. A., Masaki, T., Franke, W. W., & Holtzer, H. (1989). Differential response of myofibrillar and cytoskeletal proteins in cells treated with phorbol myristate acetate. Journal of Cell Biology, 108(3), 1079-1091.

Differential response of myofibrillar and cytoskeletal proteins in cells treated with phorbol myristate acetate. / Lin, Z.; Eshleman, James; Grund, C.; Fischman, D. A.; Masaki, T.; Franke, W. W.; Holtzer, H.

In: Journal of Cell Biology, Vol. 108, No. 3, 1989, p. 1079-1091.

Research output: Contribution to journalArticle

Lin, Z, Eshleman, J, Grund, C, Fischman, DA, Masaki, T, Franke, WW & Holtzer, H 1989, 'Differential response of myofibrillar and cytoskeletal proteins in cells treated with phorbol myristate acetate', Journal of Cell Biology, vol. 108, no. 3, pp. 1079-1091.
Lin, Z. ; Eshleman, James ; Grund, C. ; Fischman, D. A. ; Masaki, T. ; Franke, W. W. ; Holtzer, H. / Differential response of myofibrillar and cytoskeletal proteins in cells treated with phorbol myristate acetate. In: Journal of Cell Biology. 1989 ; Vol. 108, No. 3. pp. 1079-1091.
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AU - Eshleman, James

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