The long term objective of the present proposal is to understand, in molecular detail, the assembly of MHC Class I products from their subunits, and the binding of peptides to MHC Class I molecules in genetically modified mice, in living cells and in cell-free systems. The proposed experiments will address the role of the MHC Class I subunits themselves and the accessory proteins required for their proper assembly. They will also address the question of how peptides are generated and delivered to the site where combination with Class I molecules takes place, and examine the specificity of their interactions with Class I molecules. Those questions will be approached with the methods of molecular biology (gene cloning; polymerase chain reaction; transgenic mice; mutant mice produced by the ES cell homologous integration technique; in vitro transcription/translation; site directed mutagenesis; transfection), cell biology and immunology (cell culture; hybridoma production; immunochemistry; cytofluorimetry; immunoelectron microscopy). Chemical synthesis will be used not only to generate peptides of defined sequence, but also to produce peptide libraries with which to investigate the specificity of MHC Class I peptide interactions. The results obtained in this project should contribute knowledge that will help provide a rational basis for manipulating the formation of specific MHC Class I-peptide combinations. this area of experimentation is relevant not only for understanding development of the T cell repertoire, but also for tumor immunology, autoimmunity and vaccine development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37AI033456-11
Application #
6488944
Study Section
Allergy and Immunology Study Section (ALY)
Program Officer
Deckhut Augustine, Alison M
Project Start
1993-01-01
Project End
2002-12-31
Budget Start
2002-01-01
Budget End
2002-12-31
Support Year
11
Fiscal Year
2002
Total Cost
$477,964
Indirect Cost
Name
Harvard University
Department
Pathology
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Tafesse, Fikadu G; Sanyal, Sumana; Ashour, Joseph et al. (2013) Intact sphingomyelin biosynthetic pathway is essential for intracellular transport of influenza virus glycoproteins. Proc Natl Acad Sci U S A 110:6406-11
Sanyal, Sumana; Ashour, Joseph; Maruyama, Takeshi et al. (2013) Type I interferon imposes a TSG101/ISG15 checkpoint at the Golgi for glycoprotein trafficking during influenza virus infection. Cell Host Microbe 14:510-21
Witte, Martin D; Cragnolini, Juan J; Dougan, Stephanie K et al. (2012) Preparation of unnatural N-to-N and C-to-C protein fusions. Proc Natl Acad Sci U S A 109:11993-8
Kirak, Oktay; Frickel, Eva-Maria; Grotenbreg, Gijsbert M et al. (2010) Transnuclear mice with predefined T cell receptor specificities against Toxoplasma gondii obtained via SCNT. Science 328:243-8
Ernst, Robert; Mueller, Britta; Ploegh, Hidde L et al. (2009) The otubain YOD1 is a deubiquitinating enzyme that associates with p97 to facilitate protein dislocation from the ER. Mol Cell 36:28-38
Vyas, Jatin M; Van der Veen, Annemarthe G; Ploegh, Hidde L (2008) The known unknowns of antigen processing and presentation. Nat Rev Immunol 8:607-18
Mueller, Britta; Lilley, Brendan N; Ploegh, Hidde L (2006) SEL1L, the homologue of yeast Hrd3p, is involved in protein dislocation from the mammalian ER. J Cell Biol 175:261-70
Tirosh, Boaz; Iwakoshi, Neal N; Glimcher, Laurie H et al. (2006) Rapid turnover of unspliced Xbp-1 as a factor that modulates the unfolded protein response. J Biol Chem 281:5852-60
Whetstine, Johnathan R; Nottke, Amanda; Lan, Fei et al. (2006) Reversal of histone lysine trimethylation by the JMJD2 family of histone demethylases. Cell 125:467-81
Tirosh, Boaz; Iwakoshi, Neal N; Lilley, Brendan N et al. (2005) Human cytomegalovirus protein US11 provokes an unfolded protein response that may facilitate the degradation of class I major histocompatibility complex products. J Virol 79:2768-79

Showing the most recent 10 out of 28 publications