Chromosome mapping of biological pathways by fluorescence-activated cell sorting and cell fusion: human interferon gamma receptor as a model system.
Human chromosome 6 encodes both the interferon gamma receptor as well as the class I major histocompatibility complex antigens, HLA-A, -B, and -C. However, the presence of chromosome 6 in somatic cell hybrids is insufficient to confer sensitivity to human interferon gamma (Hu-IFN-gamma) as assayed by class I HLA induction; it is necessary for both human chromosomes 6 and 21 to reside in the hybrid to generate a response to Hu-IFN-gamma. Treatment of such a hamster-human hybrid, Q72-18, with Hu-IFN-gamma induces the class I HLA antigens. Q72-18 cells selected by fluorescence-activated cell sorting for the loss of class I HLA induction also lost human chromosome 21. Fusions of such cells to a hybrid that contains only human chromosome 21 reconstitutes HLA antigen induction by Hu-IFN-gamma. Furthermore, fusions of hybrids containing a translocated human chromosome 6q and the HLA-B7 gene to a line containing only human chromosome 21 or a translocated 21q also reconstitutes HLA-B7 mRNA and antigen induction by Hu-IFN-gamma. Thus the segregation of cells on the basis of a biological effect by fluorescence-activated cell sorting and reconstitution by hybrid fusion provides a strategy by which some biological pathways can be mapped at a chromosomal level.
Jung
V
V
Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854-5635.
Jones
C
C
Rashidbaigi
A
A
Geyer
D D
DD
Morse
H G
HG
Wright
R B
RB
Pestka
S
S
D015152:Blotting, Northern; D002459:Cell Fusion; D002469:Cell Separation / Q000379:methods; D002874:Chromosome Mapping / Q000379:methods*; D002891:Chromosomes, Human, Pair 21; D002896:Chromosomes, Human, Pair 6; D005434:Flow Cytometry / Q000379:methods; D006680:HLA Antigens / Q000235:genetics; D006801:Humans; D006822:Hybrid Cells; D012333:RNA, Messenger / Q000235:genetics; D011971:Receptors, Immunologic / Q000235:genetics*; D017471:Receptors, Interferon
D006680:HLA Antigens; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D017471:Receptors, Interferon