A 3 kb sequence from the mouse cellular retinoic-acid-binding protein gene upstream region mediates spatial and temporal LacZ expression in transgenic mouse embryos

Development ◽  
1991 ◽  
Vol 112 (3) ◽  
pp. 847-854
Author(s):  
L.N. Wei ◽  
G.J. Chen ◽  
Y.S. Chu ◽  
J.L. Tsao ◽  
M.C. Nguyen-Huu
Keyword(s):  
Retinoic Acid ◽  
Transgenic Mouse ◽  
Fusion Gene ◽  
Binding Protein ◽  
Initiation Site ◽  
Mouse Embryos ◽  
Upstream Region ◽  
Specific Expression ◽  
Transcriptional Initiation ◽  
Acid Binding Protein

A 3233 base pair (bp) sequence of the 5′-flanking region of the mouse cellular retinoic-acid-binding protein (CRABP) gene is determined. From this region, a 3 kb fragment located 150 bp upstream from the transcriptional initiation site is isolated and fused to a LacZ reporter sequence. Transgenic mouse embryos of this fusion gene show spatially and temporally specific expression of LacZ protein and the expression of this fusion gene at the RNA level is confirmed by RNAase protection assays, which detect specific fusion transcripts in RNA samples from tissues of transgenic mouse embryos. In contrast, transgenic mouse embryos of a shorter fusion gene containing only 583 bp from the same upstream region of the mouse CRABP gene fused to the same reporter sequence show no LacZ activities. Thus, it is concluded that the 3 kb sequence, but not the 583 bp sequence, of the mouse CRABP gene contains information for its temporally and spatially specific expression in mouse embryos.

scholarly journals THE EXPRESSION OF CELLULAR RETINOIC ACID BINDING PROTEIN (CRABP) IN DEVELOPING HEART AND SKELETAL MUSCLES OF MOUSE EMBRYOS

1991 ◽  
Vol 12 (2) ◽  
pp. 105-112 ◽  
Author(s):  
KOU ICHIHASHI ◽  
MARIKO MOMOI ◽  
TAKANORI YAMAGATA ◽  
MASAYOSHI YANAGISAWA ◽  
TAKASHI MOMOI
Keyword(s):  
Retinoic Acid ◽  
Skeletal Muscles ◽  
Binding Protein ◽  
Mouse Embryos ◽  
Acid Binding Protein ◽  
Developing Heart

The transfer of transthyretin and receptor-binding properties from the plasma retinol-binding protein to the epididymal retinoic acid-binding protein

2002 ◽  
Vol 362 (2) ◽  
pp. 265-271 ◽  
Author(s):  
Manickavasagam SUNDARAM ◽  
Daan M. F. van AALTEN ◽  
John B. C. FINDLAY ◽  
Asipu SIVAPRASADARAO
Keyword(s):  
Retinoic Acid ◽  
Binding Protein ◽  
Binding Pocket ◽  
Retinol Binding Protein ◽  
Cell Surface Receptor ◽  
Acid Binding Protein ◽  
The Family ◽  
Surface Receptor ◽  
A Cell ◽  
Plasma Retinol

Members of the lipocalin superfamily share a common structural fold, but differ from each other with respect to the molecules with which they interact. They all contain eight β-strands (A—H) that fold to form a well-defined β-barrel, which harbours a binding pocket for hydrophobic ligands. These strands are connected by loops that vary in size and structure and make up the closed and open ends of the pocket. In addition to binding ligands, some members of the family interact with other macromolecules, the specificity of which is thought to be associated with the variable loop regions. Here, we have investigated whether the macromolecular-recognition properties can be transferred from one member of the family to another. For this, we chose the prototypical lipocalin, the plasma retinol-binding protein (RBP) and its close structural homologue the epididymal retinoic acid-binding protein (ERABP). RBP exhibits three molecular-recognition properties: it binds to retinol, to transthyretin (TTR) and to a cell-surface receptor. ERABP binds retinoic acid, but whether it interacts with other macromolecules is not known. Here, we show that ERABP does not bind to TTR and the RBP receptor, but when the loops of RBP near the open end of the pocket (L-1, L-2 and L-3, connecting β-strands A—B, C—D and E—F, respectively) were substituted into the corresponding regions of ERABP, the resulting chimaera acquired the ability to bind TTR and the receptor. L-2 and L-3 were found to be the major determinants of the receptor- and TTR-binding specificities respectively. Thus we demonstrate that lipocalins serve as excellent scaffolds for engineering novel biological functions.

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