Lps(d)/Ran of endotoxin-resistant C3H/HeJ mice is defective in mediating lipopolysaccharide endotoxin responses.
C3H/HeJ inbred mice are defective in that they are highly resistant to endotoxic shock as compared with normal responder mice. Their B cells and macrophages do not respond significantly when exposed to lipopolysaccharide (LPS), whereas cells from the responder mice do. Using a functional assay, we previously isolated a cDNA, which encodes for Ran/TC4 GTPase. We now show that this gene is mutated in C3H/HeJ mice, which accounts for their resistance to endotoxin stimulation. Sequence analysis of independent mutant Lps(d)/Ran cDNAs isolated from splenic B cells of C3H/HeJ mice reveals a consistent single base substitution at position 870, where a thymidine is replaced with a cytidine. In situ hybridization maps the Lps(d)/Ran cDNA to mouse chromosome 4. By retroviral gene transfer, the wild-type Lps(n)/Ran cDNA but not the mutant Lps(d)/Ran cDNA can restore LPS responsiveness of C3H/HeJ cells. Adenoviral gene transfer in vivo with the mutant Lps(d)/Ran cDNA but not the wild-type Lps(n)/Ran cDNA rescues endotoxin-sensitive mice from septic shock. Thus Lps/Ran is an important target for LPS-mediated signal transduction, and the Lps(d)/Ran gene may be useful as a therapeutic sequence in gene therapy for endotoxemia and septic shock.
Wong
P M
PM
Department of Pathology, Fels Institute, Philadelphia, PA 19140, USA. pmcwong@compuserve.com
Kang
A
A
Chen
H
H
Yuan
Q
Q
Fan
P
P
Sultzer
B M
BM
Kan
Y W
YW
Chung
S W
SW
D020413:3' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D008070:Lipopolysaccharides / Q000633:toxicity*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics*; D012772:Shock, Septic / Q000628:therapy; D020931:ran GTP-Binding Protein
D020413:3' Untranslated Regions; D008070:Lipopolysaccharides; D009687:Nuclear Proteins; D020931:ran GTP-Binding Protein