Abstract
TGN38 and TGN41 are isoforms of a monotopic integral membrane protein which recycles between the trans Golgi network (TGN) and the cell surface, but which, at steady state, is predominantly located in the TGN. Full‐length and truncated versions of rat TGN38/41 have been expressed in monkey (COS) and human (Heb7a) cells under the control of the heavy metal inducible Metallothionein IIA promoter. This has allowed the regulated expression of TGN38/41 protein constructs to different levels in the transfected cells. These studies show that (i) controlled overexpression of TGN38/41 results in mislocalisation to parts of the endocytic pathway, (ii) a truncated version of TGN38/41, lacking the cytoplasmic domain, remains in the TGN, and (iii) there is a direct or indirect interaction between the cytoplasmic domain of TGN38/41 and γ‐adaptin.
Keywords: trans-Golgi network; Retention; γ-Adaptin; FACS, fluorescence activated cell sorter; BFA, Brefeldin A; FITC, fluorescein isothiocyanate; MTOC, Microtubule Organising Centre; TRITC, Texas red isothiocyanate; TGN, trans-Golgi network
Reaves Barbara and Banting George(1994), Overexpression of TGN38/41 leads to mislocalisation of γ-adaptin, FEBS Letters, 351, doi: 10.1016/0014-5793(94)00813-2
References
- 1. Reaves B., Banting G., J. Cell Biol., 116, (1992), 85– 94. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2. Chege N.W., Pfeffer S.R., J. Cell Biol., 11, (1990), 893– 899. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3. Luzio J.P., Brake B., Banting G., Howell K.E., Braghetta P., Stanley K.K., Biochem. J., 270, (1990), 97– 102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. Reaves B., Wilde A., Banting G., Biochem J., 283, (1992), 313– 316. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5. Ladinsky M.S., Howell K.E., Eur. J. Cell Biol., 59, (1992), 92– 105. [PubMed] [Google Scholar]
- 6. Reaves B., Horn M., Banting G., Mol. Biol. Cell., 4, (1993), 93– 105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7. Humphrey J.S., Peters P.J., Yuan L.C., Bonifacino J.S., J. Cell Biol., 120, (1993), 1123– 1135. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8. Bos K., Wraight C., Stanley K.K., EMBO J., 12, (1993), 2219– 2228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9. Wong S.H., Hong W., J. Biol. Chem., 268, (1993), 22853– 22862. [PubMed] [Google Scholar]
- 10. Ponnambalam S., Rabouille C., Luzio J.P., Nilsson T., Warren G., J. Cell Biol., 125, (1994), 253– 268. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11. Luzio J.P., Banting G., Trends Biochem., 18, (1993), 395– 398. [DOI] [PubMed] [Google Scholar]
- 12. Wilde A., Dempsey C., Banting G., J. Biol. Chem., 269, (1994), 7131– 7136. [PubMed] [Google Scholar]
- 13. Fraker P.J., Speck J.C., Biochem. Biophys. Res. Commun., 80, (1978), 849– 857. [DOI] [PubMed] [Google Scholar]
- 14. Wilde A., Reaves B., Banting G., FEBS Lett., 313, (1992), 235– 238. [DOI] [PubMed] [Google Scholar]
- 15. Horn M., Banting G., Biochem. J., 301, (1994), 69– 73. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. Ahle S., Mann A., Eichelsbacher U., Ungewickel E., EMBO J., 7, (1988), 919– 929. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17. Sambrook J., Fritsch E.F., Maniatis T., 2nd edn. Molecular Cloning: A Laboratory Manual (1989), Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY: [Google Scholar]
- 18. Gluzman Y., Cell, 23, (1981), 175– 182. [DOI] [PubMed] [Google Scholar]
- 19. Wallace D.C., Burn C.L., Eisenstadt J.M., J. Cell Biol., 67, (1979), 174– 188. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20. Chu G., Berg P., Mol. Biol. Med., 4, (1987), 277– 290. [PubMed] [Google Scholar]
- 21. Bradford M.M., Anal. Biochem., 72, (1976), 248– 254. [DOI] [PubMed] [Google Scholar]
- 22. Laemmli E.K., Nature, 227, (1970), 680– 685. [DOI] [PubMed] [Google Scholar]
- 23. Dickerson I.M., Peden K.W.C., Mains R.E., Mol. Cell Endocrinol., 64, (1989), 205– 212. [DOI] [PubMed] [Google Scholar]
- 24. Robinson M.S., Kreis T.E., Cell, 69, (1992), 129– 138. [DOI] [PubMed] [Google Scholar]
- 25. Lippincott-Schwartz J., Glickman J., Donaldson J.G., Robbins J., Kreis T.E., Seamon K.B., Sheetz M.P., Klausner R.D., J. Cell Biol., 112, (1991), 567– 577. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26. Jones S.M., Crosby J.R., Salamero J., Howell K.E., J. Cell Biol., 122, (1993), 775– 788. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 27. Piper R.C., Tai C., Slot J.W., Hahn C.S., Rice C., Huang H., James D.E., J. Cell Biol., 117, (1992), 729– 743. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 28. Asano R., Takata K., Katagiri H., Tsukuda K., Lin J.-L., Ishihara H., Inukai K., Hirano H., Yazaki Y., Oka Y., J. Biol. Chem., 267, (1992), 19636– 19641. [PubMed] [Google Scholar]
- 29. James D.E., Piper R.C., J. Cell Sci., 104, (1993), 607– 612. [DOI] [PubMed] [Google Scholar]
- 30. Nilsson R., Slusarewicz P., Hoe M.H., Warren G., FEBS Lett., 330, (1993), 1– 4. [DOI] [PubMed] [Google Scholar]
- 31. Bretscher M.S., Munro S., Science, 261, (1993), 1280– 1281. [DOI] [PubMed] [Google Scholar]
- 32. Machamer D.E., Grim M.G., Esquela A., Chung S.W., Rolls M., Ryan K., Swift A.M., Mol. Biol. Cell., 4, (1993), 695– 704. [DOI] [PMC free article] [PubMed] [Google Scholar]