Abstract
Normal fibroblasts of the vole displayed moderately spread or flattened, spindle-shaped, or polygonal morphologies and attached firmly to a substrate. Topographic features of these cells included sparse microvilli, ruffles, and filopodia. Microfilament bundles, intermediate filaments, and long microtubules generally parallel to each other, and the long axis of the cell or its extensions were present in the cytoplasm. Fibronectin was abundant, and fibronectin fibrils often formed junctions at the cell membrane with microfilament bundles. Transformation with avian sarcoma virus converted 90% of the cells to spheres 5 to 10 microns in diameter. In contrast to the normal vole cells, microfilament bundles were absent, microtubules were short and randomly arranged, and fibronectin was no longer visible. Exposure to dibutyryl cyclic AMP and testololactone caused a majority of the spherical cells to stretch and flatten, a process referred to as reverse transformation. Microtubules radiated out to the cell periphery and became parallel in cell extensions, while long microfilament bundles appeared in the cytoplasm. Parallel intermediate filaments were arranged throughout the cell. This ultrastructural analysis of reverse transformation in avian sarcoma virus-transformed vole cells detailed the status of the cytoskeletal system and showed agreement with earlier findings (Puck et al., J. Cell. Physiol. 107:399-412, 1981) using indirect immunofluorescence.
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- Asch B. B., Medina D., Brinkley B. R. Microtubules and actin-containing filaments of normal, preneoplastic, and neoplastic mouse mammary epithelial cells. Cancer Res. 1979 Mar;39(3):893–907. [PubMed] [Google Scholar]
- Ash J. F., Vogt P. K., Singer S. J. Reversion from transformed to normal phenotype by inhibition of protein synthesis in rat kidney cells infected with a temperature-sensitive mutant of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3603–3607. doi: 10.1073/pnas.73.10.3603. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bader J. P. Temperature-dependent transformation of cells infected with a mutant of Bryan Rous sarcoma virus. J Virol. 1972 Aug;10(2):267–276. doi: 10.1128/jvi.10.2.267-276.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bretscher A., Weber K. Villin is a major protein of the microvillus cytoskeleton which binds both G and F actin in a calcium-dependent manner. Cell. 1980 Jul;20(3):839–847. doi: 10.1016/0092-8674(80)90330-x. [DOI] [PubMed] [Google Scholar]
- Brinkley B. R., Fuller E. M., Highfield D. P. Cytoplasmic microtubules in normal and transformed cells in culture: analysis by tubulin antibody immunofluorescence. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4981–4985. doi: 10.1073/pnas.72.12.4981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brugge J. S., Erikson R. L. Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature. 1977 Sep 22;269(5626):346–348. doi: 10.1038/269346a0. [DOI] [PubMed] [Google Scholar]
- Chen Y. C., Hayman M. J., Vogt P. K. Properties of mammalian cells transformed by temperature-sensitive mutants of avian sarcoma virus. Cell. 1977 Jul;11(3):513–521. doi: 10.1016/0092-8674(77)90069-1. [DOI] [PubMed] [Google Scholar]
- De Mey J., Joniau M., De Brabander M., Moens W., Geuens G. Evidence for unaltered structure and in vivo assembly of microtubules in transformed cells. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1339–1343. doi: 10.1073/pnas.75.3.1339. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edelman G. M., Yahara I. Temperature-sensitive changes in surface modulating assemblies of fibroblasts transformed by mutants of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2047–2051. doi: 10.1073/pnas.73.6.2047. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. doi: 10.1016/0092-8674(79)90368-4. [DOI] [PubMed] [Google Scholar]
- Goldman R. D., Chojnacki B., Yerna M. J. Ultrastructure of microfilament bundles in baby hamster kidney (BHK-21) cells. The use of tannic acid. J Cell Biol. 1979 Mar;80(3):759–766. doi: 10.1083/jcb.80.3.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldman R. D., Yerna M. J., Schloss J. A. Localization and organization of microfilaments and related proteins in normal and virus-transformed cells. J Supramol Struct. 1976;5(2):155–183. doi: 10.1002/jss.400050206. [DOI] [PubMed] [Google Scholar]
- Haugen A., Laerum O. D. Transmission electron microscopy of fetal rat brain cells during neoplastic transformation in cell culture. J Natl Cancer Inst. 1979 Aug;63(2):455–464. [PubMed] [Google Scholar]
- Hsie A. W., Jones C., Puck T. T. Further changes in differentiation state accompanying the conversion of Chinese hamster cells of fibroblastic form by dibutyryl adenosine cyclic 3':5'-monophosphate and hormones. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1648–1652. doi: 10.1073/pnas.68.7.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsie A. W., Puck T. T. Morphological transformation of Chinese hamster cells by dibutyryl adenosine cyclic 3':5'-monophosphate and testosterone. Proc Natl Acad Sci U S A. 1971 Feb;68(2):358–361. doi: 10.1073/pnas.68.2.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsle A. W., Kawashima K., O'Neill J. P., Schröder C. H. Possible role of adenosine cyclic 3':5'-monophosphate phosphodiesterase in the morphological transformation of Chinese hamster ovary cells mediated by N6,O2-dibutyryl adenosine cyclic 3':5"-monophosphate. J Biol Chem. 1975 Feb 10;250(3):984–989. [PubMed] [Google Scholar]
- Hynes R. O., Destree A. T. 10 nm filaments in normal and transformed cells. Cell. 1978 Jan;13(1):151–163. doi: 10.1016/0092-8674(78)90146-0. [DOI] [PubMed] [Google Scholar]
- Hynes R. O., Destree A. T. Relationships between fibronectin (LETS protein) and actin. Cell. 1978 Nov;15(3):875–886. doi: 10.1016/0092-8674(78)90272-6. [DOI] [PubMed] [Google Scholar]
- Kanje M., Walum E., Edstróm A. Effects of dibutyryl cyclic AMP and cytochalasin B on cultured human glioma cells. Z Mikrosk Anat Forsch. 1979;93(3):487–496. [PubMed] [Google Scholar]
- Krebs E. G., Beavo J. A. Phosphorylation-dephosphorylation of enzymes. Annu Rev Biochem. 1979;48:923–959. doi: 10.1146/annurev.bi.48.070179.004423. [DOI] [PubMed] [Google Scholar]
- Luftig R. B., McMillan P. N., Weatherbee J. A., Weihing R. R. Increased visualization of microtubules by an improved fixation procedure. J Histochem Cytochem. 1977 Mar;25(3):175–187. doi: 10.1177/25.3.402414. [DOI] [PubMed] [Google Scholar]
- McClain D. A., Maness P. F., Edelman G. M. Assay for early cytoplasmic effects of the src gene product of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2750–2754. doi: 10.1073/pnas.75.6.2750. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meek W. D., Porter K. R., Puck T. T. The ultrastructure of process formation following treatment with db-cAMP of a Chinese hamster ovary x Chinese hamster brain cell hybrid. Exp Cell Res. 1980 Apr;126(2):359–374. doi: 10.1016/0014-4827(80)90275-x. [DOI] [PubMed] [Google Scholar]
- Meek W. D., Puck T. T. Role of the microfibrillar system in knob action of transformed cells. J Supramol Struct. 1979;12(3):335–354. doi: 10.1002/jss.400120306. [DOI] [PubMed] [Google Scholar]
- Nielson S. E., Puck T. T. Deposition of fibronectin in the course of reverse transformation of Chinese hamster ovary cells by cyclic AMP. Proc Natl Acad Sci U S A. 1980 Feb;77(2):985–989. doi: 10.1073/pnas.77.2.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osborn M., Weber K. Simian virus 40 gene A function and maintenance of transformation. J Virol. 1975 Mar;15(3):636–644. doi: 10.1128/jvi.15.3.636-644.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osborn M., Weber K. The display of microtubules in transformed cells. Cell. 1977 Nov;12(3):561–571. doi: 10.1016/0092-8674(77)90257-4. [DOI] [PubMed] [Google Scholar]
- Podlubnaya Z. A., Tskhovrebova L. A., Zaalishtsbvili M. M., Stefanenko G. A. Electron microscopic study of alpha-actinin. J Mol Biol. 1975 Feb 25;92(2):357–359. doi: 10.1016/0022-2836(75)90234-x. [DOI] [PubMed] [Google Scholar]
- Pollack R., Osborn M., Weber K. Patterns of organization of actin and myosin in normal and transformed cultured cells. Proc Natl Acad Sci U S A. 1975 Mar;72(3):994–998. doi: 10.1073/pnas.72.3.994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Porter K. R., Puck T. T., Hsie A. W., Kelley D. An electron microscopy study of the effects on dibutyryl cyclic AMP on Chinese hamster ovary cells. Cell. 1974 Jul;2(3):145–162. doi: 10.1016/0092-8674(74)90089-0. [DOI] [PubMed] [Google Scholar]
- Puck T. T. Cyclic AMP, the microtubule-microfilament system, and cancer. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4491–4495. doi: 10.1073/pnas.74.10.4491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Puck T. T., Erikson R. L., Meek W. D., Nielson S. E. Reverse transformation of vole cells transformed by avian sarcoma virus containing the src gene. J Cell Physiol. 1981 Jun;107(3):399–412. doi: 10.1002/jcp.1041070312. [DOI] [PubMed] [Google Scholar]
- Puck T. T., Waldren C. A., Hsie A. W. Membrane dynamics in the action of dibutyryl adenosine 3':5'-cyclic monophosphate and testosterone on mammalian cells. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1943–1947. doi: 10.1073/pnas.69.7.1943. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Purchio A. F., Erikson E., Erikson R. L. Translation of 35S and of subgenomic regions of avian sarcoma virus RNA. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4661–4665. doi: 10.1073/pnas.74.10.4661. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rohrschneider L. R. Adhesion plaques of Rous sarcoma virus-transformed cells contain the src gene product. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3514–3518. doi: 10.1073/pnas.77.6.3514. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sen A., Todaro G. J. A murine sarcoma virus-associated protein kinase: interaction with actin and microtubular protein. Cell. 1979 Jun;17(2):347–356. doi: 10.1016/0092-8674(79)90161-2. [DOI] [PubMed] [Google Scholar]
- Singer I. I. The fibronexus: a transmembrane association of fibronectin-containing fibers and bundles of 5 nm microfilaments in hamster and human fibroblasts. Cell. 1979 Mar;16(3):675–685. doi: 10.1016/0092-8674(79)90040-0. [DOI] [PubMed] [Google Scholar]
- Singer S. J., Ash J. F., Bourguignon L. Y., Heggeness M. H., Louvard D. Transmembrane interactions and the mechanisms of transport of proteins across membranes. J Supramol Struct. 1978;9(3):373–389. doi: 10.1002/jss.400090308. [DOI] [PubMed] [Google Scholar]
- Storrie B., Puck T. T., Wenger L. The role of butyrate in the reverse transformation reaction in mammalian cells. J Cell Physiol. 1978 Jan;94(1):69–75. doi: 10.1002/jcp.1040940109. [DOI] [PubMed] [Google Scholar]
- Vollet J. J., Brugge J. S., Noonan C. A., Butel J. S. The role of SV40 gene A in the alteration of microfilaments in transformed cells. Exp Cell Res. 1977 Mar 1;105(1):119–126. doi: 10.1016/0014-4827(77)90157-4. [DOI] [PubMed] [Google Scholar]
- Wang E., Goldberg A. R. Changes in microfilament organization and surface topogrophy upon transformation of chick embryo fibroblasts with Rous sarcoma virus. Proc Natl Acad Sci U S A. 1976 Nov;73(11):4065–4069. doi: 10.1073/pnas.73.11.4065. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Willingham M. C., Jay G., Pastan I. Localization of the ASV src gene product to the plasma membrane of transformed cells by electron microscopic immunocytochemistry. Cell. 1979 Sep;18(1):125–134. doi: 10.1016/0092-8674(79)90361-1. [DOI] [PubMed] [Google Scholar]
- Willingham M. C., Pastan I. Cyclic amp and cell morphology in cultured fibroblasts. Effects on cell shape, microfilament and microtubule distribution, and orientation to substratum. J Cell Biol. 1975 Oct;67(1):146–159. doi: 10.1083/jcb.67.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Willingham M. C., Pastan I., Shih T. Y., Scolnick E. M. Localization of the src gene product of the Harvey strain of MSV to plasma membrane of transformed cells by electron microscopic immunocytochemistry. Cell. 1980 Apr;19(4):1005–1014. doi: 10.1016/0092-8674(80)90091-4. [DOI] [PubMed] [Google Scholar]
- Wulf E., Deboben A., Bautz F. A., Faulstich H., Wieland T. Fluorescent phallotoxin, a tool for the visualization of cellular actin. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4498–4502. doi: 10.1073/pnas.76.9.4498. [DOI] [PMC free article] [PubMed] [Google Scholar]











