Novel alternatively spliced exon in the extracellular ligand-binding domain of the pituitary adenylate cyclase-activating polypeptide (PACAP) type 1 receptor (PAC1R) selectively increases ligand affinity and alters signal transduction coupling during spermatogenesis
Pb. Daniel et al., Novel alternatively spliced exon in the extracellular ligand-binding domain of the pituitary adenylate cyclase-activating polypeptide (PACAP) type 1 receptor (PAC1R) selectively increases ligand affinity and alters signal transduction coupling during spermatogenesis, J BIOL CHEM, 276(16), 2001, pp. 12938-12944
The expression of the paracrine signaling hormone pituitary adenylate cycla
se-activating polypeptide (PACAP) is regulated in a cyclical fashion during
the Ig-day spermatogenic cycle of the adult rat testis. The precise functi
ons of PACAP in the development of germ cells are uncertain, but cycle- and
stage-specific expression may augment cAMP-regulated gene expression in ge
rm cells and associated Sertoli cells. Here we report the existence of a he
retofore unrecognized exon in the extracellular domain of the PACAP type 1
receptor (PAC1R) that is alternatively spliced during the spermatogenic cyc
le in the rat testis, This splice variant encodes a full-length receptor wi
th the insertion of an additional 72 base pairs encoding 24 amino acids (ex
on 3a) between coding exons 3 and 4. The PAC1R(3a) mRNA is preferentially d
etected in seminiferous tubules and is expressed at the highest levels in r
ound spermatids and Sertoli cells. Analyses of ligand binding and signaling
functions in stably transfected HEK293 cells expressing the two receptor i
soforms reveals a 6-fold increase in the affinity of the PAC1R(3a) to bind
PACAP-38, and alterations in its coupling to both cAMP and inositol phospha
te signaling pathways relative to the wild type PAC1R. These findings sugge
st that the extracellular region between coding exons 3 and g of PAC1R may
play an important role in the regulation of the relative ligand affinities
and the relative coupling to G, (cAMP) and G, (inositol phosphates) signal
transduction pathways during spermatogenesis.