title: The C2 cytosolic loop of adenylyl cyclase interacts with the activated form of G alpha s.
abstract: Using the yeast two-hybrid system, we studied the physical interaction between the complete C1 and C2 cytosolic domains of Xenopus laevis type 9 (xl9C1, xl9C2) and the C2 domain of rat type 6 (r6C2) adenylyl cyclase (AC). Heterodimerization between xl9C1 and xl9C2 and homodimerization between C2 (but not C1) domains was observed. Interaction between C2 and human G alpha s (hG alpha s) was also detected and was dependent on G alpha s activation. In contrast X. laevis G alpha s (xlG alpha s), which is 92% identical to hG alpha s, was unable to interact with any of the three AC cytosolic domains tested, corroborating previous findings that showed no effector activation. Through the construction of chimeras, we demonstrated that the amino-terminal half of xlG alpha s was responsible for the lack of interaction with AC. Chimeras between mouse G alpha i2 and G alpha s (N-mG alpha i2/C-G alpha s), that have previously shown to activate AC to a higher extent than wild-type G alpha s, also interacted with the C2 cytosolic domain and with a higher affinity. Interestingly, N-mG alpha i2/C-xlG alpha s chimera was not only able to interact with C2 but also with the C1 cytosolic domain.
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Biológicas, Universidad de Concepción, Chile.
authors.forename: M
authors.initials: M
authors.lastname: Torrejón
authors.forename: A M
authors.initials: AM
authors.lastname: Genevière
authors.forename: V
authors.initials: V
authors.lastname: Echeverría
authors.forename: L
authors.initials: L
authors.lastname: Guzmán
authors.forename: M V
authors.initials: MV
authors.lastname: Hinrichs
authors.forename: J
authors.initials: J
authors.lastname: Olate
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014982:Xenopus laevis; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D001616:beta-Galactosidase; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases