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. 1995 Jul;69(7):4095–4102. doi: 10.1128/jvi.69.7.4095-4102.1995

Membrane permeabilization by different regions of the human immunodeficiency virus type 1 transmembrane glycoprotein gp41.

J Arroyo 1, M Boceta 1, M E González 1, M Michel 1, L Carrasco 1
PMCID: PMC189144  PMID: 7769667

Abstract

The transmembrane glycoprotein (gp41) of human immunodeficiency virus type 1 (HIV-1) has been implicated in the cytopathology observed during HIV infection. The first amino acids located at the amino terminus are involved in membrane fusion and syncytium formation, while sequences located at the carboxy terminus have been predicted to interact with membranes and modify membrane permeability. The HIV-1 gp41 gene has been cloned and expressed in Escherichia coli cells by using pET vectors to analyze changes in membrane permeability produced by this protein. This system is well suited for expressing toxic genes in an inducible manner and for analyzing the function of proteins that modify membrane permeability. gp41 enhances the permeability of the bacterial membrane to hygromycin B despite the low level of expression of this protein. To localize the regions of gp41 responsible for these effects, a number of fragments spanning different portions of gp41 were inducibly expressed in E. coli. Two regions of gp41 were shown to increase membrane permeability: one located at the carboxy terminus, where two highly amphipathic helices have been predicted, and another one corresponding to the membrane-spanning domain. Expression of the central region of gp41 comprising this domain was highly lytic for E. coli cells and increased membrane permeability to a number of compounds. These findings are discussed in the light of HIV-induced cytopathology and gp41 structure.

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Selected References

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  1. Alonso M. A., Carrasco L. Action of membrane-active compounds on mammalian cells. Permeabilization of human cells by ionophores to inhibitors of translation and transcription. Eur J Biochem. 1980 Aug;109(2):535–540. doi: 10.1111/j.1432-1033.1980.tb04825.x. [DOI] [PubMed] [Google Scholar]
  2. Antoni B. A., Stein S. B., Rabson A. B. Regulation of human immunodeficiency virus infection: implications for pathogenesis. Adv Virus Res. 1994;43:53–145. doi: 10.1016/s0065-3527(08)60047-0. [DOI] [PubMed] [Google Scholar]
  3. Bergeron L., Sullivan N., Sodroski J. Target cell-specific determinants of membrane fusion within the human immunodeficiency virus type 1 gp120 third variable region and gp41 amino terminus. J Virol. 1992 Apr;66(4):2389–2397. doi: 10.1128/jvi.66.4.2389-2397.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bernheimer A. W., Rudy B. Interactions between membranes and cytolytic peptides. Biochim Biophys Acta. 1986 Jun 12;864(1):123–141. doi: 10.1016/0304-4157(86)90018-3. [DOI] [PubMed] [Google Scholar]
  5. Bosch M. L., Earl P. L., Fargnoli K., Picciafuoco S., Giombini F., Wong-Staal F., Franchini G. Identification of the fusion peptide of primate immunodeficiency viruses. Science. 1989 May 12;244(4905):694–697. doi: 10.1126/science.2541505. [DOI] [PubMed] [Google Scholar]
  6. Cao J., Bergeron L., Helseth E., Thali M., Repke H., Sodroski J. Effects of amino acid changes in the extracellular domain of the human immunodeficiency virus type 1 gp41 envelope glycoprotein. J Virol. 1993 May;67(5):2747–2755. doi: 10.1128/jvi.67.5.2747-2755.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cao J., Vasir B., Sodroski J. G. Changes in the cytopathic effects of human immunodeficiency virus type 1 associated with a single amino acid alteration in the ectodomain of the gp41 transmembrane glycoprotein. J Virol. 1994 Jul;68(7):4662–4668. doi: 10.1128/jvi.68.7.4662-4668.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Carrasco L. Membrane leakiness after viral infection and a new approach to the development of antiviral agents. Nature. 1978 Apr 20;272(5655):694–699. doi: 10.1038/272694a0. [DOI] [PubMed] [Google Scholar]
  9. Carrasco L. Modification of membrane permeability by animal viruses. Adv Virus Res. 1995;45:61–112. doi: 10.1016/S0065-3527(08)60058-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Carrasco L., Otero M. J., Castrillo J. L. Modification of membrane permeability by animal viruses. Pharmacol Ther. 1989;40(2):171–212. doi: 10.1016/0163-7258(89)90096-x. [DOI] [PubMed] [Google Scholar]
  11. Chakrabarti L., Emerman M., Tiollais P., Sonigo P. The cytoplasmic domain of simian immunodeficiency virus transmembrane protein modulates infectivity. J Virol. 1989 Oct;63(10):4395–4403. doi: 10.1128/jvi.63.10.4395-4403.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chen S. S. Functional role of the zipper motif region of human immunodeficiency virus type 1 transmembrane protein gp41. J Virol. 1994 Mar;68(3):2002–2010. doi: 10.1128/jvi.68.3.2002-2010.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chen S. S., Lee C. N., Lee W. R., McIntosh K., Lee T. H. Mutational analysis of the leucine zipper-like motif of the human immunodeficiency virus type 1 envelope transmembrane glycoprotein. J Virol. 1993 Jun;67(6):3615–3619. doi: 10.1128/jvi.67.6.3615-3619.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Cloyd M. W., Lynn W. S. Perturbation of host-cell membrane is a primary mechanism of HIV cytopathology. Virology. 1991 Apr;181(2):500–511. doi: 10.1016/0042-6822(91)90882-c. [DOI] [PubMed] [Google Scholar]
  15. Contreras A., Carrasco L. Selective inhibition of protein synthesis in virus-infected mammalian cells. J Virol. 1979 Jan;29(1):114–122. doi: 10.1128/jvi.29.1.114-122.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dedera D., Gu R. L., Ratner L. Conserved cysteine residues in the human immunodeficiency virus type 1 transmembrane envelope protein are essential for precursor envelope cleavage. J Virol. 1992 Feb;66(2):1207–1209. doi: 10.1128/jvi.66.2.1207-1209.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dempsey C. E. The actions of melittin on membranes. Biochim Biophys Acta. 1990 May 7;1031(2):143–161. doi: 10.1016/0304-4157(90)90006-x. [DOI] [PubMed] [Google Scholar]
  18. Dubay J. W., Roberts S. J., Hahn B. H., Hunter E. Truncation of the human immunodeficiency virus type 1 transmembrane glycoprotein cytoplasmic domain blocks virus infectivity. J Virol. 1992 Nov;66(11):6616–6625. doi: 10.1128/jvi.66.11.6616-6625.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Eisenberg D., Wesson M. The most highly amphiphilic alpha-helices include two amino acid segments in human immunodeficiency virus glycoprotein 41. Biopolymers. 1990 Jan;29(1):171–177. doi: 10.1002/bip.360290122. [DOI] [PubMed] [Google Scholar]
  20. Fauci A. S. Multifactorial nature of human immunodeficiency virus disease: implications for therapy. Science. 1993 Nov 12;262(5136):1011–1018. doi: 10.1126/science.8235617. [DOI] [PubMed] [Google Scholar]
  21. Fisher A. G., Ratner L., Mitsuya H., Marselle L. M., Harper M. E., Broder S., Gallo R. C., Wong-Staal F. Infectious mutants of HTLV-III with changes in the 3' region and markedly reduced cytopathic effects. Science. 1986 Aug 8;233(4764):655–659. doi: 10.1126/science.3014663. [DOI] [PubMed] [Google Scholar]
  22. Freed E. O., Myers D. J., Risser R. Characterization of the fusion domain of the human immunodeficiency virus type 1 envelope glycoprotein gp41. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4650–4654. doi: 10.1073/pnas.87.12.4650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gabuzda D. H., Lever A., Terwilliger E., Sodroski J. Effects of deletions in the cytoplasmic domain on biological functions of human immunodeficiency virus type 1 envelope glycoproteins. J Virol. 1992 Jun;66(6):3306–3315. doi: 10.1128/jvi.66.6.3306-3315.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Gallaher W. R. Detection of a fusion peptide sequence in the transmembrane protein of human immunodeficiency virus. Cell. 1987 Jul 31;50(3):327–328. doi: 10.1016/0092-8674(87)90485-5. [DOI] [PubMed] [Google Scholar]
  25. Garry R. F., Gottlieb A. A., Zuckerman K. P., Pace J. R., Frank T. W., Bostick D. A. Cell surface effects of human immunodeficiency virus. Biosci Rep. 1988 Feb;8(1):35–48. doi: 10.1007/BF01128970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Gawrisch K., Han K. H., Yang J. S., Bergelson L. D., Ferretti J. A. Interaction of peptide fragment 828-848 of the envelope glycoprotein of human immunodeficiency virus type I with lipid bilayers. Biochemistry. 1993 Mar 30;32(12):3112–3118. doi: 10.1021/bi00063a024. [DOI] [PubMed] [Google Scholar]
  27. Guinea R., Carrasco L. Influenza virus M2 protein modifies membrane permeability in E. coli cells. FEBS Lett. 1994 May 2;343(3):242–246. doi: 10.1016/0014-5793(94)80564-4. [DOI] [PubMed] [Google Scholar]
  28. Helseth E., Olshevsky U., Furman C., Sodroski J. Human immunodeficiency virus type 1 gp120 envelope glycoprotein regions important for association with the gp41 transmembrane glycoprotein. J Virol. 1991 Apr;65(4):2119–2123. doi: 10.1128/jvi.65.4.2119-2123.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kell D. B., Ryder H. M., Kaprelyants A. S., Westerhoff H. V. Quantifying heterogeneity: flow cytometry of bacterial cultures. Antonie Van Leeuwenhoek. 1991 Oct-Nov;60(3-4):145–158. doi: 10.1007/BF00430362. [DOI] [PubMed] [Google Scholar]
  30. Kowalski M., Bergeron L., Dorfman T., Haseltine W., Sodroski J. Attenuation of human immunodeficiency virus type 1 cytopathic effect by a mutation affecting the transmembrane envelope glycoprotein. J Virol. 1991 Jan;65(1):281–291. doi: 10.1128/jvi.65.1.281-291.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lama J., Carrasco L. Expression of poliovirus nonstructural proteins in Escherichia coli cells. Modification of membrane permeability induced by 2B and 3A. J Biol Chem. 1992 Aug 5;267(22):15932–15937. [PubMed] [Google Scholar]
  32. Lama J., Carrasco L. Inducible expression of a toxic poliovirus membrane protein in Escherichia coli: comparative studies using different expression systems based on T7 promoters. Biochem Biophys Res Commun. 1992 Nov 16;188(3):972–981. doi: 10.1016/0006-291x(92)91327-m. [DOI] [PubMed] [Google Scholar]
  33. Lama J., Guinea R., Martinez-Abarca F., Carrasco L. Cloning and inducible synthesis of poliovirus nonstructural proteins. Gene. 1992 Aug 15;117(2):185–192. doi: 10.1016/0378-1119(92)90728-8. [DOI] [PubMed] [Google Scholar]
  34. Lee S. J., Hu W., Fisher A. G., Looney D. J., Kao V. F., Mitsuya H., Ratner L., Wong-Staal F. Role of the carboxy-terminal portion of the HIV-1 transmembrane protein in viral transmission and cytopathogenicity. AIDS Res Hum Retroviruses. 1989 Aug;5(4):441–449. doi: 10.1089/aid.1989.5.441. [DOI] [PubMed] [Google Scholar]
  35. Levy J. A. Pathogenesis of human immunodeficiency virus infection. Microbiol Rev. 1993 Mar;57(1):183–289. doi: 10.1128/mr.57.1.183-289.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lynn W. S., Tweedale A., Cloyd M. W. Human immunodeficiency virus (HIV-1) cytotoxicity: perturbation of the cell membrane and depression of phospholipid synthesis. Virology. 1988 Mar;163(1):43–51. doi: 10.1016/0042-6822(88)90232-2. [DOI] [PubMed] [Google Scholar]
  37. Miller M. A., Cloyd M. W., Liebmann J., Rinaldo C. R., Jr, Islam K. R., Wang S. Z., Mietzner T. A., Montelaro R. C. Alterations in cell membrane permeability by the lentivirus lytic peptide (LLP-1) of HIV-1 transmembrane protein. Virology. 1993 Sep;196(1):89–100. doi: 10.1006/viro.1993.1457. [DOI] [PubMed] [Google Scholar]
  38. Miller M. A., Garry R. F., Jaynes J. M., Montelaro R. C. A structural correlation between lentivirus transmembrane proteins and natural cytolytic peptides. AIDS Res Hum Retroviruses. 1991 Jun;7(6):511–519. doi: 10.1089/aid.1991.7.511. [DOI] [PubMed] [Google Scholar]
  39. Nieva J. L., Nir S., Muga A., Goñi F. M., Wilschut J. Interaction of the HIV-1 fusion peptide with phospholipid vesicles: different structural requirements for fusion and leakage. Biochemistry. 1994 Mar 22;33(11):3201–3209. doi: 10.1021/bi00177a009. [DOI] [PubMed] [Google Scholar]
  40. Owens R. J., Burke C., Rose J. K. Mutations in the membrane-spanning domain of the human immunodeficiency virus envelope glycoprotein that affect fusion activity. J Virol. 1994 Jan;68(1):570–574. doi: 10.1128/jvi.68.1.570-574.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Perez L. G., O'Donnell M. A., Stephens E. B. The transmembrane glycoprotein of human immunodeficiency virus type 1 induces syncytium formation in the absence of the receptor binding glycoprotein. J Virol. 1992 Jul;66(7):4134–4143. doi: 10.1128/jvi.66.7.4134-4143.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Pinter A., Honnen W. J., Tilley S. A., Bona C., Zaghouani H., Gorny M. K., Zolla-Pazner S. Oligomeric structure of gp41, the transmembrane protein of human immunodeficiency virus type 1. J Virol. 1989 Jun;63(6):2674–2679. doi: 10.1128/jvi.63.6.2674-2679.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Popovic M., Sarin P. S., Robert-Gurroff M., Kalyanaraman V. S., Mann D., Minowada J., Gallo R. C. Isolation and transmission of human retrovirus (human t-cell leukemia virus). Science. 1983 Feb 18;219(4586):856–859. doi: 10.1126/science.6600519. [DOI] [PubMed] [Google Scholar]
  44. Ritter G. D., Jr, Mulligan M. J., Lydy S. L., Compans R. W. Cell fusion activity of the simian immunodeficiency virus envelope protein is modulated by the intracytoplasmic domain. Virology. 1993 Nov;197(1):255–264. doi: 10.1006/viro.1993.1586. [DOI] [PubMed] [Google Scholar]
  45. Sanz M. A., Pérez L., Carrasco L. Semliki Forest virus 6K protein modifies membrane permeability after inducible expression in Escherichia coli cells. J Biol Chem. 1994 Apr 22;269(16):12106–12110. [PubMed] [Google Scholar]
  46. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  47. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  48. Studier F. W. Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system. J Mol Biol. 1991 May 5;219(1):37–44. doi: 10.1016/0022-2836(91)90855-z. [DOI] [PubMed] [Google Scholar]
  49. Syu W. J., Lee W. R., Du B., Yu Q. C., Essex M., Lee T. H. Role of conserved gp41 cysteine residues in the processing of human immunodeficiency virus envelope precursor and viral infectivity. J Virol. 1991 Nov;65(11):6349–6352. doi: 10.1128/jvi.65.11.6349-6352.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Thomas D. J., Wall J. S., Hainfeld J. F., Kaczorek M., Booy F. P., Trus B. L., Eiserling F. A., Steven A. C. gp160, the envelope glycoprotein of human immunodeficiency virus type 1, is a dimer of 125-kilodalton subunits stabilized through interactions between their gp41 domains. J Virol. 1991 Jul;65(7):3797–3803. doi: 10.1128/jvi.65.7.3797-3803.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Venable R. M., Pastor R. W., Brooks B. R., Carson F. W. Theoretically determined three-dimensional structures for amphipathic segments of the HIV-1 gp41 envelope protein. AIDS Res Hum Retroviruses. 1989 Feb;5(1):7–22. doi: 10.1089/aid.1989.5.7. [DOI] [PubMed] [Google Scholar]
  52. Weiss R. A. How does HIV cause AIDS? Science. 1993 May 28;260(5112):1273–1279. doi: 10.1126/science.8493571. [DOI] [PubMed] [Google Scholar]
  53. de la Fuente J. M., Alvarez A., Nombela C., Sanchez M. Flow cytometric analysis of Saccharomyces cerevisiae autolytic mutants and protoplasts. Yeast. 1992 Jan;8(1):39–45. doi: 10.1002/yea.320080104. [DOI] [PubMed] [Google Scholar]

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