MOLECULAR-CLONING AND EXPRESSION ANALYSIS OF RAT RGS12 AND RGS14

We report the cloning of two novel rat regulators of G-protein signali ng (RGS) cDNAs using a degenerate PCR strategy. The rRgs12 and rRgs14 cDNAs encode predicted polypeptides of 1387 and 544 amino acids, respe ctively, We have also identified the human orthologue of rRgs12 by ali gnment of cosmid sequences in the database which map the human RGS12 g ene to chromosome 4p16.3. Furthermore, we identified human ESTs with h igh homology to rRgs14 which map to human chromosome 5qter. Northern b lot analysis indicates that rRgs14 is expressed at high levels in brai n, lung, and spleen, whereas rRgs12 is expressed at high levels in bra in and lung and lower levels in testis, heart, and spleen. Analysis of the predicted rRGS12 and rRGS14 polypeptides indicates that they are closely related and possess regions of homology outside of the conserv ed RGS domain. We have also identified conserved regions in RGS12 whic h are similar to protein domains found in mouse rhophilin and coiled-c oil proteins suggesting possible interactions with ras-like G-proteins . (C) 1997 Academic Press.