Major histocompatibility complex class I genes in murine fibrosarcoma IC9 are down regulated at the level of the chromatin structure.
The fibrosarcoma IC9 is deficient in the expression of the major histocompatibility complex class I genes Kb, Kk, and Dk and expresses only the Db molecule. Because class I deficiency may enable tumor cells to escape the immune response by cytotoxic T lymphocytes, we investigated why the class I genes are not expressed. Expression of the silent class I genes could not be induced, but all known DNA-binding factors specific for class I genes could be detected in nuclear extracts of IC9 cells. After cloning of the silent Kb gene from the IC9 cells and subsequent transfection of this cloned Kb gene into LTK- and IC9 cells, normal Kb antigens were expressed on the cell surface of both cell lines. Digestion of the chromatin of IC9 cells with micrococcal nuclease and DNase I showed a decreased nuclease sensitivity of the silent class I genes in comparison with active genes and the absence of DNase I hypersensitive sites in the promoter region of the silent Dk gene. These findings demonstrate that class I expression is turned off by a cis-acting regulatory mechanism at the level of the chromatin structure.
Maschek
U
U
Institute of Immunology and Genetics, German Cancer Research Center, Heidelberg, Federal Republic of Germany.
Pülm
W
W
Segal
S
S
Hämmerling
G J
GJ
D000483:Alleles; D000818:Animals; D000942:Antigens, Bacterial; D000954:Antigens, Surface / Q000235:genetics; D015139:Blotting, Southern; D002843:Chromatin / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I; D005354:Fibrosarcoma / Q000235:genetics*; D005786:Gene Expression Regulation; D005805:Genes, MHC Class I; D006822:Hybrid Cells; D007371:Interferon-gamma / Q000494:pharmacology; D051379:Mice; D009154:Mutation; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D002843:Chromatin; C030659:K antigens; D012333:RNA, Messenger; D014157:Transcription Factors; D007371:Interferon-gamma; D004247:DNA; D003850:Deoxyribonuclease I