Molecular cloning and expression of an adenylyl cyclase from Xenopus laevis oocytes.
We have cloned a cDNA that encodes a novel Xenopus laevis oocyte adenylyl cyclase (xlAC) using oligonucleotides against conserved mammalian adenylyl cyclase regions. The isolated cDNA is 4372 bp long with an open reading frame of 4065 nucleotides which encodes a protein of 1355 amino acids. Comparison of the deduced amino acid sequence with previously cloned mammalian adenylyl cyclases shows a low identity, 19.7% with type 2 rat adenylyl cyclase and 24.2% with type 4 rat adenylyl cyclase, indicating that this Xenopus isoform represents a new member of this protein family. Gene expression studies of the xlAC by reverse PCR showed that this gene is expressed in all oogenesis stages but not during early embryogenesis. Expression of the xlAC in COS-7 cells resulted in increased basal AC activity, that was stimulated by forskolin, Gpp(NH)p and aluminium fluoride, and was insensitive to calcium and calcium-calmodulin (Ca2(+)-CaM).
Torrejón
M
M
Departamento de Fisiopatología, Facultad de Ciencias Biológicas, Universidad de Concepción, Chile.
Echeverría
V
V
Retamales
G
G
Herrera
L
L
Hinrichs
M V
MV
Olate
J
J
D000262:Adenylyl Cyclases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009865:Oocytes / Q000737:chemistry / Q000201:enzymology* / Q000254:growth & development; D009866:Oogenesis / Q000235:genetics; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection; D014982:Xenopus laevis
D018076:DNA, Complementary; D012333:RNA, Messenger; D000262:Adenylyl Cyclases