Cloning, characterization and expression of a functional mouse ACTH receptor.
A polymerase chain reaction-generated mouse ACTH-R probe was used to screen a mouse genomic library, and a clone of 13kb containing the entire coding sequence was isolated. The coding sequence shows 84% homology with the human gene at the DNA level and encodes a peptide with 89% homology to the human ACTH-R. This gene is expressed as a major transcript of 1.8kb in the mouse adrenal gland. The gene was expressed in HeLa cells and cAMP production in response to either ACTH or alpha-MSH was measured. cAMP increased in an ACTH dose dependent manner suggesting an EC50 of 7 x 10(-10)M ACTH. alpha-MSH was without effect on this receptor. In conclusion we have cloned a mouse ACTH receptor gene and demonstrated for the first time its expression and functional effect in HeLa cells.
Cammas
F M
FM
Department of Chemical Endocrinology, St Bartholomew's Hospital Medical College, London, United Kingdom.
Kapas
S
S
Barker
S
S
Clark
A J
AJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000096:biosynthesis; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D015698:Genomic Library; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018043:Receptors, Corticotropin / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014162:Transfection
D018043:Receptors, Corticotropin; D004247:DNA; D000242:Cyclic AMP