An aspartate insulin receptor chimera mitogenically activates fibroblasts

被引:9
作者
Biemann, HP
Harmer, SL
Koshland, DE
机构
[1] UNIV CALIF BERKELEY,DEPT MOL & CELLULAR BIOL,BERKELEY,CA 94720
[2] GENZYME CORP,DEPT CELL BIOL,CAMBRIDGE,MA 02139
[3] UNIV CALIF SAN FRANCISCO,GEORGE WILLIAMS HOOPER FDN,SAN FRANCISCO,CA 94143
关键词
D O I
10.1074/jbc.271.44.27927
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A gene encoding the ligand-binding domain of the Escherichia coli aspartate receptor fused to the cytoplasmic domain of the insulin receptor tyrosine kinase to produce the chimeric aspartate insulin receptor (AIR) was expressed in mammalian cells. A murine fibroblast transfectant line designated CA3 was generated that stably expressed the AIR receptor. This 70,000 M(r) receptor containing the tyrosine kinase of the insulin receptor was recognized by aspartate receptor-specific antisera. When isolated in cellular membrane preparations, AIR was found to be capable of autophosphorylation and phosphorylation of histone H2B on tyrosine. The receptor was found to be predominately cytoplasmic and to be situated in the endoplasmic reticulum and Golgi membranes by immunofluorescence imaging of CA3 cells. Mitogenic effects of AIR were observed; CA3 cells continued DNA synthesis under serum deprivation conditions that prevented parental cells from cycling. These results demonstrate that a chimeric receptor containing procaryotic transmembrane sequences is expressed by a eucaryotic cell in intracellular membranes and functionally couples to cellular signaling pathways.
引用
收藏
页码:27927 / 27930
页数:4
相关论文
共 29 条
[1]   ASPARTATE RECEPTORS OF ESCHERICHIA-COLI AND SALMONELLA-TYPHIMURIUM BIND LIGAND WITH NEGATIVE AND HALF-OF-THE-SITES COOPERATIVITY [J].
BIEMANN, HP ;
KOSHLAND, DE .
BIOCHEMISTRY, 1994, 33 (03) :629-634
[2]   SOLUBILIZATION OF A VECTORIAL TRANSMEMBRANE RECEPTOR IN FUNCTIONAL FORM - ASPARTATE RECEPTOR OF CHEMOTAXIS [J].
BOGONEZ, E ;
KOSHLAND, DE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (15) :4891-4895
[3]  
CELIS JE, 1994, CELL BIOL LAB HDB, V1, P297
[4]   ATTRACTANT-INDUCED AND DISULFIDE-INDUCED CONFORMATIONAL-CHANGES IN THE LIGAND-BINDING DOMAIN OF THE CHEMOTAXIS ASPARTATE RECEPTOR - A F-19 NMR-STUDY [J].
DANIELSON, MA ;
BIEMANN, HP ;
KOSHLAND, DE ;
FALKE, JJ .
BIOCHEMISTRY, 1994, 33 (20) :6100-6109
[5]   SAFE AND EFFICIENT GENERATION OF RECOMBINANT RETROVIRUSES WITH AMPHOTROPIC AND ECOTROPIC HOST RANGES [J].
DANOS, O ;
MULLIGAN, RC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (17) :6460-6464
[6]  
DEMEYTS P, 1994, DIABETOLOGIA, V37, P135
[7]  
DINTZIS SM, 1994, J BIOL CHEM, V269, P12159
[8]   A MEMBRANE-ANCHORED CYTOPLASMIC DOMAIN OF THE HUMAN INSULIN-RECEPTOR MEDIATES A CONSTITUTIVELY ELEVATED INSULIN-INDEPENDENT UPTAKE OF 2-DEOXYGLUCOSE [J].
ELLIS, L ;
MORGAN, DO ;
CLAUSER, E ;
ROTH, RA ;
RUTTER, WJ .
MOLECULAR ENDOCRINOLOGY, 1987, 1 (01) :15-24
[9]   LINKING FUNCTIONAL DOMAINS OF THE HUMAN INSULIN-RECEPTOR WITH THE BACTERIAL ASPARTATE RECEPTOR [J].
ELLIS, L ;
MORGAN, DO ;
KOSHLAND, DE ;
CLAUSER, E ;
MOE, GR ;
BOLLAG, G ;
ROTH, RA ;
RUTTER, WJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (21) :8137-8141
[10]  
HARLOW E, 1988, ANTIBODIES LABORATOR, P313