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Identification through ENU genome‐wide mutagenesis of a novel mutant mouse that lacks MHC‐II as a result of missense mutation in SFX‐5
Author(s) -
Chen YiTing,
Chang MeiLing,
Tan SihHui,
Kung John
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.417
Subject(s) - biology , genetics , mutant , gene , phenotype , missense mutation , mutagenesis , allele , mutation , genome , microbiology and biotechnology
With the human genome sequenced, the race is on to understand the functions of the 30,000 or so genes. Ethyl‐nitrosourea (ENU) was used to treat C57BL/6 (B6) male mice, followed by a 3‐generation breeding scheme to generate G3 mice that were screened for immunological phenotypes under recessive genetic control. The primary phenotype of one mutant, P‐334, was originally identified by a total lack of IA expression on PBL of ENU‐mutagenized G3 mice. Recessive mode of inheritance was confirmed by mating an affected G3 male to wildtype B6 females to generate F2 offspring. No affected phenotype was found in F1 mice and approximately 1 in 4 F2 mice were pheno‐deviants. Real‐time RT‐PCR revealed depressed expression of both I‐Aα and I‐Aβ mRNA. Since the CIITA protein complex is known to regulate all I‐A gene expression, DNA sequencing of genes encoding CIITA and its associated proteins was performed. A single base (T to A) mutation resulted in a change of Phe at position 134 to Ser. No NP‐specific antibody response was observed in P‐334 mutant mice immunized with NP‐CGG. P‐334 mutant mice carry a novel and non‐functional SFX‐5 allele and may be used to define domain structures of importance in SFX‐5 interaction with other proteins that make up the CIITA master regulator complex.