Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1985 Nov 1;101(5):1713–1723. doi: 10.1083/jcb.101.5.1713

Blocked coated pits in AtT20 cells result from endocytosis of budding retrovirions

PMCID: PMC2113951  PMID: 2865267

Abstract

AtT20 cells support the replication of two endogenous retroviruses, a murine leukemia virus and a mouse mammary tumor virus. On glass or plastic substrates, AtT20 cells grow in clumps. In this situation, retroviruses budding from the plasma membrane of one cell can, on rare occasions, be invested by coated pits in the plasma membranes of contiguous cells. These pits can invaginate to depths of 2,000-4,000 A within the cytoplasm drawing with them the viral buds which remain connected to their parental cells by tubular stalks, some of which are only 225 +/- 15 A in diameter. These stalks run down the straight necks of the pits from the buds to the parental cell surfaces. Several lines of evidence indicate that these unique structures are blocked such that neither endocytosis nor budding can go to completion, and that they persist for several hours. The properties of these blocked coated pits are relevant to models of both endocytosis and viral budding. First, they indicate that the invagination of a coated pit is not absolutely dependent on its pinching off to form a coated vesicle, but that uncoating appears to be dependent upon the generation of a free vesicle. Secondly, they suggest that the final stages in the maturation of a retroviral core into a mature nucleoid are dependent on the detachment of the bud from its parental cell and that the driving force of budding is the association of viral transmembrane proteins with viral core proteins. An explanation is offered to account for the formation of these structures despite the phenomenon of viral interference.

Full Text

The Full Text of this article is available as a PDF (2.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andersen K. B., Nexø B. A. Entry of murine retrovirus into mouse fibroblasts. Virology. 1983 Feb;125(1):85–98. doi: 10.1016/0042-6822(83)90065-x. [DOI] [PubMed] [Google Scholar]
  2. Armbruster B. L., Carlemalm E., Chiovetti R., Garavito R. M., Hobot J. A., Kellenberger E., Villiger W. Specimen preparation for electron microscopy using low temperature embedding resins. J Microsc. 1982 Apr;126(Pt 1):77–85. doi: 10.1111/j.1365-2818.1982.tb00358.x. [DOI] [PubMed] [Google Scholar]
  3. Birdwell C. R., Strauss E. G., Strauss J. H. Replication of Sindbis virus. 3. An electron microscopic study of virus maturation using the surface replica technique. Virology. 1973 Dec;56(2):429–438. doi: 10.1016/0042-6822(73)90047-0. [DOI] [PubMed] [Google Scholar]
  4. Dales S., Hanafusa H. Penetration and intracellular release of the genomes of avian RNA tumor viruses. Virology. 1972 Nov;50(2):440–458. doi: 10.1016/0042-6822(72)90396-0. [DOI] [PubMed] [Google Scholar]
  5. Danon D., Goldstein L., Marikovsky Y., Skutelsky E. Use of cationized ferritin as a label of negative charges on cell surfaces. J Ultrastruct Res. 1972 Mar;38(5):500–510. doi: 10.1016/0022-5320(72)90087-1. [DOI] [PubMed] [Google Scholar]
  6. Dickson C., Atterwill M. Polyproteins related to the major core protein of mouse mammary tumor virus. J Virol. 1978 Jun;26(3):660–672. doi: 10.1128/jvi.26.3.660-672.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Farquhar M. G. Recovery of surface membrane in anterior pituitary cells. Variations in traffic detected with anionic and cationic ferritin. J Cell Biol. 1978 Jun;77(3):R35–R42. doi: 10.1083/jcb.77.3.r35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fine D., Schochetman G. Type D primate retroviruses: a review. Cancer Res. 1978 Oct;38(10):3123–3139. [PubMed] [Google Scholar]
  9. Griffiths G., Simons K., Warren G., Tokuyasu K. T. Immunoelectron microscopy using thin, frozen sections: application to studies of the intracellular transport of Semliki Forest virus spike glycoproteins. Methods Enzymol. 1983;96:466–485. doi: 10.1016/s0076-6879(83)96041-x. [DOI] [PubMed] [Google Scholar]
  10. Gumbiner B., Kelly R. B. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell. 1982 Jan;28(1):51–59. doi: 10.1016/0092-8674(82)90374-9. [DOI] [PubMed] [Google Scholar]
  11. Louvard D., Morris C., Warren G., Stanley K., Winkler F., Reggio H. A monoclonal antibody to the heavy chain of clathrin. EMBO J. 1983;2(10):1655–1664. doi: 10.1002/j.1460-2075.1983.tb01640.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Marsh M., Helenius A. Adsorptive endocytosis of Semliki Forest virus. J Mol Biol. 1980 Sep 25;142(3):439–454. doi: 10.1016/0022-2836(80)90281-8. [DOI] [PubMed] [Google Scholar]
  13. Mooren H. W., Prins F. A., Herbrink P., Warnaar S. O. Electron microscopic studies on the role of the envelope antigens of R-MuLV-ts29 in budding. Virology. 1981 Aug;113(1):254–262. doi: 10.1016/0042-6822(81)90152-5. [DOI] [PubMed] [Google Scholar]
  14. Pastan I. H., Willingham M. C. Journey to the center of the cell: role of the receptosome. Science. 1981 Oct 30;214(4520):504–509. doi: 10.1126/science.6170111. [DOI] [PubMed] [Google Scholar]
  15. Perry M. M., Gilbert A. B. Yolk transport in the ovarian follicle of the hen (Gallus domesticus): lipoprotein-like particles at the periphery of the oocyte in the rapid growth phase. J Cell Sci. 1979 Oct;39:257–272. doi: 10.1242/jcs.39.1.257. [DOI] [PubMed] [Google Scholar]
  16. Petersen O. W., van Deurs B. Serial-section analysis of coated pits and vesicles involved in adsorptive pinocytosis in cultured fibroblasts. J Cell Biol. 1983 Jan;96(1):277–281. doi: 10.1083/jcb.96.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schmid S. L., Braell W. A., Schlossman D. M., Rothman J. E. A role for clathrin light chains in the recognition of clathrin cages by 'uncoating ATPase'. Nature. 1984 Sep 20;311(5983):228–231. doi: 10.1038/311228a0. [DOI] [PubMed] [Google Scholar]
  18. Simons K., Warren G. Semliki Forest virus: a probe for membrane traffic in the animal cell. Adv Protein Chem. 1984;36:79–132. doi: 10.1016/S0065-3233(08)60296-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tooze J., Tooze S. A. Infection of AtT20 murine pituitary tumour cells by mouse hepatitis virus strain A59: virus budding is restricted to the Golgi region. Eur J Cell Biol. 1985 May;37:203–212. [PubMed] [Google Scholar]
  20. Wall D. A., Wilson G., Hubbard A. L. The galactose-specific recognition system of mammalian liver: the route of ligand internalization in rat hepatocytes. Cell. 1980 Aug;21(1):79–93. doi: 10.1016/0092-8674(80)90116-6. [DOI] [PubMed] [Google Scholar]
  21. Weihing R. R. Cytochalasin B inhibits actin-related gelation of HeLa cell extracts. J Cell Biol. 1976 Oct;71(1):303–307. doi: 10.1083/jcb.71.1.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Willingham M. C., Rutherford A. V., Gallo M. G., Wehland J., Dickson R. B., Schlegel R., Pastan I. H. Receptor-mediated endocytosis in cultured fibroblasts: cryptic coated pits and the formation of receptosomes. J Histochem Cytochem. 1981 Sep;29(9):1003–1013. doi: 10.1177/29.9.6169759. [DOI] [PubMed] [Google Scholar]
  23. van Deurs B., Petersen O. W., Bundgaard M. Identification of free coated pinocytic vesicles in Swiss 3T3 cells. EMBO J. 1984 Sep;3(9):1959–1964. doi: 10.1002/j.1460-2075.1984.tb02076.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES