Tk. Hinson et al., IDENTIFICATION OF PUTATIVE TRANSMEMBRANE RECEPTOR SEQUENCES HOMOLOGOUS TO THE CALCIUM-SENSING G-PROTEIN-COUPLED RECEPTOR, Genomics, 45(2), 1997, pp. 279-289
The sensing of extracellular calcium is a general paradigm for regulat
ing diverse cellular functions in many tissues. A calcium-sensing rece
ptor (Casr) belonging to the metabotropic glutamate family of G-protei
n-coupled receptors (GPCR) that transduces the effects of extracellula
r calcium in the parathyroid gland as well as other tissues has been i
dentified. The diversity of GPCR families and the recent finding of ca
lcium sensing in cells lacking the known Casr suggest the existence of
additional receptors related to Casr. By polymerase chain reaction (P
CR) amplification and screening of genomic libraries, we have identifi
ed multiple Casr-related sequences (Casr-rs) in the mouse. Using prime
rs designed to regions of the first and third intracellular loops of C
asr, we initially PCR amplified a 497-bp Casr-related sequence (Casr-r
s1) with high homology to Casr. The deduced protein sequence of Casr-r
s1 is 63% similar and 40% identical to Casr over the available transme
mbrane region. We screened a mouse genomic library with a Casr-rs1 pro
be and identified two additional Casr-related sequences (Casr-rs2 and
Casr-rs3). In the predicted transmembrane domain, Casr-rs2 and Casr-rs
3 are 95% identical to Casr-rs1. We mapped Casr-rs1 to mouse Chromosom
e (Chr) 7 by interspecific backcross analysis, whereas the known Casr
localizes to mouse Chr 16. By fluorescence in situ hybridization, Casr
-rs2 also localized to mouse Chr 7 and Casr-rs3 mapped to mouse Chr 4.
We were able to distinguish Casr-rs1 from Casr-rs2 by PCR using speci
fic primers, suggesting that they are distinct genes clustered on Chr
7. By RT-PCR, we identified additional Casr-rs transcripts in mouse ki
dney, brain, testis, embryo, and MC3T3-E1 osteoblasts, but not in lung
or liver. The homologous sequence in mouse kidney, embryo, and MC3T3-
E1 osteoblasts, designated Casr-rs4 has a deduced amino acid sequence
that is 100% similar and 97% identical to that of Casr-rs1. The sequen
ce amplified from mouse brain, Casr-rs5, has a deduced protein sequenc
e that is 96% similar and 92% identical to that of Casr-rs1. Our findi
ngs establish the existence of a novel multimembered family of Casr-re
lated sequences in the mouse which may encode receptors that transduce
responses to diverse extracellular cations. (C) 1997 Academic Press.