V. Asghari et al., DOPAMINE D4 RECEPTOR REPEAT - ANALYSIS OF DIFFERENT NATIVE AND MUTANTFORMS OF THE HUMAN AND RAT GENES, Molecular pharmacology, 46(2), 1994, pp. 364-373
Recent molecular characterization of the human D4 gene has revealed th
e existence of various polymorphic forms of this receptor. These varia
tions are found in the putative third cytoplasmic loop region and enco
de a variable number of repeats of 16 amino acids in length. In the pr
esent study we have compared the pharmacological binding profiles of s
even different polymorphic variants of the human D4 receptor, the rat
D4 receptor, and two different human D4 receptor mutants that were del
eted in the repeat sequence. For this purpose we cloned the rat D4 rec
eptor gene and compared its gene structure and its pharmacological bin
ding profile with those of the D4.4 and D4.7 genes. The rat and human
D4 genes display a high degree of sequence similarity, especially in t
he coding regions. An Alu repeat sequence was identified in the first
intron of the human D4 gene but is not present in the rat D4 gene. Fur
thermore, using the polymerase chain reaction we cloned 3-, 5-, 6-, an
d 9-fold repeat sequences. These cloned repeat sequences were used for
the reconstruction of full length cDNAs encoding D4.3, D4.5, D4.6, an
d D4.9, respectively. These novel forms of the human D4 receptor, as w
ell as the previously cloned D4.2, D4.4, and D4.7 forms, were transien
tly expressed in COS-7 cells. All of the different forms of the human
and rat D4 receptors and repeat deletion mutants displayed similar bin
ding profiles for all ligands tested, although small differences were
observed. The affinity for dopamine could be decreased by guanosine-5'
-(beta,gamma-imido)triphosphate with the different forms of the D4 rec
eptor, including the two receptor mutants that were deleted in the rep
eat sequence. These data suggest that the polymorphic repeat sequence
has little influence on D4 binding profiles and might not be essential
for G protein interaction.