Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1994 Sep 1;180(3):1107–1113. doi: 10.1084/jem.180.3.1107

The CLIP region of invariant chain plays a critical role in regulating major histocompatibility complex class II folding, transport, and peptide occupancy

PMCID: PMC2191661  PMID: 8064228

Abstract

Invariant chain (Ii) contributes in a number of distinct ways to the proper functioning of major histocompatibility complex (MHC) class II molecules. These include promoting effective association and folding of newly synthesized MHC class II alpha and beta subunits, increasing transit of assembled heterodimers out of the endoplasmic reticulum (ER), inhibiting class II peptide binding, and facilitating class II movement to or accumulation in endosomes/lysosomes. Although the cytoplasmic tail of Ii makes a key contribution to the endocytic localization of class II, the relationship between the structure of Ii and its other diverse functions remains unknown. We show here that two thirds of the lumenal segment of Ii can be eliminated without affecting its contributions to the secretory pathway events of class II folding, ER to Golgi transport, or inhibition of peptide binding. These same experiments reveal that a short (25 residue) contiguous internal segment of Ii (the CLIP region), frequently found associated with purified MHC class II molecules, is critical for all three functions. Together with other recent findings, these results raise the possibility that the contributions of Ii to the early postsynthetic behavior of class II may depend on its interaction with the class II binding site. This would be consistent with the intracellular behavior of unoccupied MHC class I and class II molecules as incompletely folded proteins and imply a related structural basis for the similar contributions of Ii to class II and of short peptides to class I assembly and transport.

Full Text

The Full Text of this article is available as a PDF (846.5 KB).

Selected References

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

  1. Anderson M. S., Miller J. Invariant chain can function as a chaperone protein for class II major histocompatibility complex molecules. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2282–2286. doi: 10.1073/pnas.89.6.2282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bakke O., Dobberstein B. MHC class II-associated invariant chain contains a sorting signal for endosomal compartments. Cell. 1990 Nov 16;63(4):707–716. doi: 10.1016/0092-8674(90)90137-4. [DOI] [PubMed] [Google Scholar]
  3. Bikoff E. K., Huang L. Y., Episkopou V., van Meerwijk J., Germain R. N., Robertson E. J. Defective major histocompatibility complex class II assembly, transport, peptide acquisition, and CD4+ T cell selection in mice lacking invariant chain expression. J Exp Med. 1993 Jun 1;177(6):1699–1712. doi: 10.1084/jem.177.6.1699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonnerot C., Marks M. S., Cosson P., Robertson E. J., Bikoff E. K., Germain R. N., Bonifacino J. S. Association with BiP and aggregation of class II MHC molecules synthesized in the absence of invariant chain. EMBO J. 1994 Feb 15;13(4):934–944. doi: 10.1002/j.1460-2075.1994.tb06338.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chicz R. M., Urban R. G., Lane W. S., Gorga J. C., Stern L. J., Vignali D. A., Strominger J. L. Predominant naturally processed peptides bound to HLA-DR1 are derived from MHC-related molecules and are heterogeneous in size. Nature. 1992 Aug 27;358(6389):764–768. doi: 10.1038/358764a0. [DOI] [PubMed] [Google Scholar]
  6. Elliott E. A., Drake J. R., Amigorena S., Elsemore J., Webster P., Mellman I., Flavell R. A. The invariant chain is required for intracellular transport and function of major histocompatibility complex class II molecules. J Exp Med. 1994 Feb 1;179(2):681–694. doi: 10.1084/jem.179.2.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fling S. P., Arp B., Pious D. HLA-DMA and -DMB genes are both required for MHC class II/peptide complex formation in antigen-presenting cells. Nature. 1994 Apr 7;368(6471):554–558. doi: 10.1038/368554a0. [DOI] [PubMed] [Google Scholar]
  8. Freisewinkel I. M., Schenck K., Koch N. The segment of invariant chain that is critical for association with major histocompatibility complex class II molecules contains the sequence of a peptide eluted from class II polypeptides. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9703–9706. doi: 10.1073/pnas.90.20.9703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Germain R. N., Hendrix L. R. MHC class II structure, occupancy and surface expression determined by post-endoplasmic reticulum antigen binding. Nature. 1991 Sep 12;353(6340):134–139. doi: 10.1038/353134a0. [DOI] [PubMed] [Google Scholar]
  10. Germain R. N. MHC-dependent antigen processing and peptide presentation: providing ligands for T lymphocyte activation. Cell. 1994 Jan 28;76(2):287–299. doi: 10.1016/0092-8674(94)90336-0. [DOI] [PubMed] [Google Scholar]
  11. Hunt D. F., Michel H., Dickinson T. A., Shabanowitz J., Cox A. L., Sakaguchi K., Appella E., Grey H. M., Sette A. Peptides presented to the immune system by the murine class II major histocompatibility complex molecule I-Ad. Science. 1992 Jun 26;256(5065):1817–1820. doi: 10.1126/science.1319610. [DOI] [PubMed] [Google Scholar]
  12. Janeway C. A., Jr, Conrad P. J., Lerner E. A., Babich J., Wettstein P., Murphy D. B. Monoclonal antibodies specific for Ia glycoproteins raised by immunization with activated T cells: possible role of T cellbound Ia antigens as targets of immunoregulatory T cells. J Immunol. 1984 Feb;132(2):662–667. [PubMed] [Google Scholar]
  13. Karp D. R., Jenkins R. N., Long E. O. Distinct binding sites on HLA-DR for invariant chain and staphylococcal enterotoxins. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9657–9661. doi: 10.1073/pnas.89.20.9657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Koch N., Koch S., Hämmerling G. J. Ia invariant chain detected on lymphocyte surfaces by monoclonal antibody. Nature. 1982 Oct 14;299(5884):644–645. doi: 10.1038/299644a0. [DOI] [PubMed] [Google Scholar]
  15. Koch N., Lauer W., Habicht J., Dobberstein B. Primary structure of the gene for the murine Ia antigen-associated invariant chains (Ii). An alternatively spliced exon encodes a cysteine-rich domain highly homologous to a repetitive sequence of thyroglobulin. EMBO J. 1987 Jun;6(6):1677–1683. doi: 10.1002/j.1460-2075.1987.tb02417.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lamb C. A., Yewdell J. W., Bennink J. R., Cresswell P. Invariant chain targets HLA class II molecules to acidic endosomes containing internalized influenza virus. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):5998–6002. doi: 10.1073/pnas.88.14.5998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Layet C., Germain R. N. Invariant chain promotes egress of poorly expressed, haplotype-mismatched class II major histocompatibility complex A alpha A beta dimers from the endoplasmic reticulum/cis-Golgi compartment. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2346–2350. doi: 10.1073/pnas.88.6.2346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lopata M. A., Cleveland D. W., Sollner-Webb B. High level transient expression of a chloramphenicol acetyl transferase gene by DEAE-dextran mediated DNA transfection coupled with a dimethyl sulfoxide or glycerol shock treatment. Nucleic Acids Res. 1984 Jul 25;12(14):5707–5717. doi: 10.1093/nar/12.14.5707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lotteau V., Teyton L., Peleraux A., Nilsson T., Karlsson L., Schmid S. L., Quaranta V., Peterson P. A. Intracellular transport of class II MHC molecules directed by invariant chain. Nature. 1990 Dec 13;348(6302):600–605. doi: 10.1038/348600a0. [DOI] [PubMed] [Google Scholar]
  20. Miller J., Germain R. N. Efficient cell surface expression of class II MHC molecules in the absence of associated invariant chain. J Exp Med. 1986 Nov 1;164(5):1478–1489. doi: 10.1084/jem.164.5.1478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Morris P., Shaman J., Attaya M., Amaya M., Goodman S., Bergman C., Monaco J. J., Mellins E. An essential role for HLA-DM in antigen presentation by class II major histocompatibility molecules. Nature. 1994 Apr 7;368(6471):551–554. doi: 10.1038/368551a0. [DOI] [PubMed] [Google Scholar]
  22. Murphy D. B., Lo D., Rath S., Brinster R. L., Flavell R. A., Slanetz A., Janeway C. A., Jr A novel MHC class II epitope expressed in thymic medulla but not cortex. Nature. 1989 Apr 27;338(6218):765–768. doi: 10.1038/338765a0. [DOI] [PubMed] [Google Scholar]
  23. Peterson M., Miller J. Antigen presentation enhanced by the alternatively spliced invariant chain gene product p41. Nature. 1992 Jun 18;357(6379):596–598. doi: 10.1038/357596a0. [DOI] [PubMed] [Google Scholar]
  24. Riberdy J. M., Newcomb J. R., Surman M. J., Barbosa J. A., Cresswell P. HLA-DR molecules from an antigen-processing mutant cell line are associated with invariant chain peptides. Nature. 1992 Dec 3;360(6403):474–477. doi: 10.1038/360474a0. [DOI] [PubMed] [Google Scholar]
  25. Roche P. A., Cresswell P. Invariant chain association with HLA-DR molecules inhibits immunogenic peptide binding. Nature. 1990 Jun 14;345(6276):615–618. doi: 10.1038/345615a0. [DOI] [PubMed] [Google Scholar]
  26. Roche P. A., Marks M. S., Cresswell P. Formation of a nine-subunit complex by HLA class II glycoproteins and the invariant chain. Nature. 1991 Dec 5;354(6352):392–394. doi: 10.1038/354392a0. [DOI] [PubMed] [Google Scholar]
  27. Romagnoli P., Layet C., Yewdell J., Bakke O., Germain R. N. Relationship between invariant chain expression and major histocompatibility complex class II transport into early and late endocytic compartments. J Exp Med. 1993 Mar 1;177(3):583–596. doi: 10.1084/jem.177.3.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rudensky AYu, Preston-Hurlburt P., Hong S. C., Barlow A., Janeway C. A., Jr Sequence analysis of peptides bound to MHC class II molecules. Nature. 1991 Oct 17;353(6345):622–627. doi: 10.1038/353622a0. [DOI] [PubMed] [Google Scholar]
  29. Sadegh-Nasseri S., Germain R. N. A role for peptide in determining MHC class II structure. Nature. 1991 Sep 12;353(6340):167–170. doi: 10.1038/353167a0. [DOI] [PubMed] [Google Scholar]
  30. Sadegh-Nasseri S., Germain R. N. How MHC class II molecules work: peptide-dependent completion of protein folding. Immunol Today. 1992 Feb;13(2):43–46. doi: 10.1016/0167-5699(92)90131-P. [DOI] [PubMed] [Google Scholar]
  31. Sette A., Adorini L., Colon S. M., Buus S., Grey H. M. Capacity of intact proteins to bind to MHC class II molecules. J Immunol. 1989 Aug 15;143(4):1265–1267. [PubMed] [Google Scholar]
  32. Sette A., Ceman S., Kubo R. T., Sakaguchi K., Appella E., Hunt D. F., Davis T. A., Michel H., Shabanowitz J., Rudersdorf R. Invariant chain peptides in most HLA-DR molecules of an antigen-processing mutant. Science. 1992 Dec 11;258(5089):1801–1804. doi: 10.1126/science.1465617. [DOI] [PubMed] [Google Scholar]
  33. Stern L. J., Brown J. H., Jardetzky T. S., Gorga J. C., Urban R. G., Strominger J. L., Wiley D. C. Crystal structure of the human class II MHC protein HLA-DR1 complexed with an influenza virus peptide. Nature. 1994 Mar 17;368(6468):215–221. doi: 10.1038/368215a0. [DOI] [PubMed] [Google Scholar]
  34. Townsend A., Elliott T., Cerundolo V., Foster L., Barber B., Tse A. Assembly of MHC class I molecules analyzed in vitro. Cell. 1990 Jul 27;62(2):285–295. doi: 10.1016/0092-8674(90)90366-m. [DOI] [PubMed] [Google Scholar]
  35. Townsend A., Ohlén C., Bastin J., Ljunggren H. G., Foster L., Kärre K. Association of class I major histocompatibility heavy and light chains induced by viral peptides. Nature. 1989 Aug 10;340(6233):443–448. doi: 10.1038/340443a0. [DOI] [PubMed] [Google Scholar]
  36. Viville S., Neefjes J., Lotteau V., Dierich A., Lemeur M., Ploegh H., Benoist C., Mathis D. Mice lacking the MHC class II-associated invariant chain. Cell. 1993 Feb 26;72(4):635–648. doi: 10.1016/0092-8674(93)90081-z. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES