Coding GNAS mutations leading to hormone resistance impair in vitro agonist- and cholera toxin-induced adenosine cyclic 3',5'-monophosphate formation mediated by human XLalphas.

Abstract

Most loss of function mutations of GNAS identified in different forms of pseudohypoparathyroidism disrupt not only the stimulatory G protein alpha-subunit (Gsalpha), but also its paternally expressed variant, XLalphas. However, the possibility that XLalphas deficiency contributes to disease pathogenesis has remained unexplored. We therefore examined the signaling property of human XLalphas and the effects of one novel (XLalphas(H704P) or Gsalpha(H362P)) and two previously described (XLalphas(DelI724) and XLalphas(Y733X) or Gsalpha(DelI382) and Gsalpha(Y391X), respectively) GNAS mutations on either XLalphas or Gsalpha activity. Confocal immunofluorescence microscopy detected human XLalphas immunoreactivity at the plasma membrane of transduced mouse embryonic fibroblasts null for endogenous Gsalpha and XLalphas (Gnas(E2-/E2-) cells). Cholera toxin- and isoproterenol-induced cAMP accumulation in Gnas(E2-/E2-) cells transiently expressing wild-type human XLalphas was similar to that in cells transiently expressing wild-type Gsalpha. Human XLalphas, like Gsalpha, mediated PTH-induced cAMP accumulation in Gnas(E2-/E2-) cells coexpressing PTH receptor type 1 and either of these proteins. Moreover, overexpression of human XLalphas or Gsalpha markedly enhanced the PTH-induced cAMP accumulation in opossum kidney cells that endogenously express PTH receptor type 1. In contrast, each XLalphas mutant failed to mediate isoproterenol- and PTH-induced cAMP accumulation in transduced Gnas(E2-/E2-) cells. XLalphas(DelI724) showed a reduced cholera toxin response over the basal level compared with wild-type XLalphas, and XLalphas(H704P) completely failed to respond to cholera toxin. These findings were comparable to those observed with each corresponding Gsalpha mutant transiently expressed in Gnas(E2-/E2-) cells. Thus, mutations that typically inactivate Gsalpha also impair XLalphas activity, consistent with a possible role for XLalphas deficiency in diseases caused by paternal GNAS mutations.

• Authors: Agnès Linglart, Matthew J Mahon, Mohammad A Kerachian, David M Berlach, Geoffrey N Hendy, Harald Jüppner, Murat Bastepe
• Journal: Endocrinology (Endocrinology) Vol. 147 Issue 5 Pg. 2253-62 (May 2006) ISSN: 0013-7227 [Print] United States
• PMID: 16484323 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Chromogranins
• DNA, Complementary
• Hormones
• Peptides
• Cholera Toxin
• Cyclic AMP
• GNAS protein, human
• Gnas protein, mouse
• GTP-Binding Protein alpha Subunits, Gs
• Isoproterenol

Topics

• Adenoviridae (genetics)
• Animals
• Base Sequence
• Blotting, Western
• Cell Membrane (metabolism)
• Cholera Toxin (chemistry, metabolism)
• Chromogranins
• Cyclic AMP (chemistry, metabolism)
• DNA Mutational Analysis
• DNA, Complementary (metabolism)
• Dose-Response Relationship, Drug
• Exons
• Female
• Fibroblasts (metabolism)
• GTP-Binding Protein alpha Subunits, Gs (genetics, physiology)
• Hormones (metabolism)
• Humans
• Immunohistochemistry
• In Vitro Techniques
• Infant
• Isoproterenol (pharmacology)
• Male
• Mice
• Microscopy, Confocal
• Microscopy, Fluorescence
• Models, Genetic
• Molecular Sequence Data
• Mutation
• Peptides (chemistry)
• Signal Transduction