Genetic selection of regulatory mutants of mammalian adenylyl cyclases.
Initial steps in the identification of the Gs alpha-binding site present in mammalian adenylyl cyclases can be achieved with the use of the yeast genetic system described. It must be stressed that this system serves as a means to identify mutants that are candidates; biochemical analysis of these mutants is a next and necessary step in the confirmation of these phenotypes. The system described can be readily adapted for the isolation of additional classes of mammalian adenylyl cyclase mutants including mutants with altered regulation toward forskolin, catalytic abnormalities, or enhanced sensitivities toward activators. In addition, this system can be employed for the isolation of constitutively active adenylyl cyclase mutants, or by coexpressing other adenylyl cyclase isoforms and their known regulators, mutations in the binding sites for these molecules can be elucidated.
Clapp
Peter
P
Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Capper
Austin B
AB
Taussig
Ronald
R
D000262:Adenylyl Cyclases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D004285:Dogs; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D015723:Gene Library; D005821:Genetic Techniques; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D009154:Mutation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic
D007527:Isoenzymes; D011994:Recombinant Proteins; D019205:GTP-Binding Protein alpha Subunits, Gs; D000262:Adenylyl Cyclases; C419510:adenylyl cyclase type V