Breeding stock-specific variation in peptidylglycine alpha-amidating monooxygenase messenger ribonucleic acid splicing in rat pituitary.
Peptidylglycine alpha-amidating monooxygenase (PAM) is a bifunctional enzyme that catalyzes the carboxyl-terminal amidation of glycine-extended peptides in a two-step reaction involving a monooxygenase and a lyase. Several forms of PAM messenger RNA result from alternative splicing of the single copy PAM gene. The presence of alternately spliced exon A between the two enzymatic domains allows endoproteolytic cleavage to occur in selected tissues, generating soluble monooxygenase and membrane lyase from integral membrane PAM. While using an exon A antiserum, we made the unexpected observation that Charles River Sprague Dawley rats expressed forms of PAM containing exon A in their pituitaries, whereas Harlan Sprague Dawley rats did not. Forms of PAM containing exon A were expressed in the atrium and hypothalamus of both types of Sprague Dawley rat, although in different proportions. PAM transmembrane domain splicing also differed between rat breeders, and full-length PAM-1 was not prevalent in the anterior pituitary of either type of rat. Despite striking differences in PAM splicing, no differences in levels of monooxygenase or lyase activity were observed in tissue or serum samples. The splicing patterns of other alternatively spliced genes, pituitary adenylate cyclase-activating polypeptide receptor type 1 and cardiac troponin T, did not vary with rat breeder. Strain-specific variations in the splicing of transcripts such as PAM must be taken into account in analyzing the resultant proteins, and knowledge of these differences should identify variations with functional significance.
Ciccotosto
G D
GD
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Hand
T A
TA
Mains
R E
RE
Eipper
B A
BA
D017398:Alternative Splicing; D000818:Animals; D001947:Breeding / Q000379:methods; D005091:Exons; D015971:Gene Expression Regulation, Enzymologic; D014644:Genetic Variation; D007031:Hypothalamus / Q000201:enzymology*; D008297:Male; D006899:Mixed Function Oxygenases / Q000235:genetics*; D009097:Multienzyme Complexes; D010902:Pituitary Gland / Q000201:enzymology*; D010903:Pituitary Gland, Anterior / Q000201:enzymology; D016133:Polymerase Chain Reaction; D011333:Pro-Opiomelanocortin / Q000096:biosynthesis / Q000235:genetics; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity
D009097:Multienzyme Complexes; D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase