A new missense mutation in the growth hormone-releasing hormone receptor gene in familial isolated GH deficiency.

2.50
Hdl Handle:
http://hdl.handle.net/10541/79013
Title:
A new missense mutation in the growth hormone-releasing hormone receptor gene in familial isolated GH deficiency.
Authors:
Carakushansky, Mauri; Whatmore, Andrew J; Clayton, Peter E; Shalet, Stephen M; Gleeson, Helena K; Price, David A; Levine, Michael A; Salvatori, Roberto
Abstract:
OBJECTIVE: Mutations in the GH-releasing hormone (GHRH) receptor (GHRHR) gene (GHRHR) cause autosomal recessive familial isolated GH deficiency (IGHD). We searched for GHRHR mutations in two siblings with IGHD type IB and a history of parental consanguinity. DESIGN: We analyzed peripheral genomic DNA of an index patient. After identifying a novel mutation in the GHRHR, we performed functional studies in order to confirm that the mutation causes receptor malfunction. METHODS: The entire GHRHR was analyzed in the index case by denaturing gradient gel electrophoresis. Abnormally migrating bands were isolated and sequenced. The mutated area was then sequenced in all family members whose DNA was available. The newly found mutation was inserted into a GHRHR cDNA. Wild-type and mutant cDNAs were expressed into CHO cells and the cyclic AMP (cAMP) response to GHRH was measured. In order to determine whether the mutant receptor was properly expressed on the cell membrane surface, CHO cells were transfected with wild-type or mutant GHRHR cDNA containing a FLAG epitope tag in the extracellular N-terminus. RESULTS: Both patients were homozygous for a new missense mutation in codon 176, corresponding to the second transmembrane domain of the receptor protein that replaces alanine with valine (A176V). The mother and three unaffected siblings were heterozygous for the mutation; DNA from the father was not available. Cells expressing the A176V receptor had a significantly reduced cAMP response to GHRH, despite appropriate expression on the cell surface. CONCLUSIONS: We describe two siblings with IGHD due to a new mutation in the GHRHR that disrupts GHRH signaling and leads to GHRH resistance.
Affiliation:
Divisions of Pediatric Endocrinology and Endocrinology and The Ilyssa Center for Molecular and Cellular Endocrinology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.
Citation:
A new missense mutation in the growth hormone-releasing hormone receptor gene in familial isolated GH deficiency. 2003, 148 (1):25-30 Eur. J. Endocrinol.
Journal:
European Journal of Endocrinology
Issue Date:
Jan-2003
URI:
http://hdl.handle.net/10541/79013
PubMed ID:
12534354
Type:
Article
Language:
en
ISSN:
0804-4643
Appears in Collections:
All Christie Publications

Full metadata record

DC FieldValue Language
dc.contributor.authorCarakushansky, Mauri-
dc.contributor.authorWhatmore, Andrew J-
dc.contributor.authorClayton, Peter E-
dc.contributor.authorShalet, Stephen M-
dc.contributor.authorGleeson, Helena K-
dc.contributor.authorPrice, David A-
dc.contributor.authorLevine, Michael A-
dc.contributor.authorSalvatori, Roberto-
dc.date.accessioned2009-08-27T16:14:04Z-
dc.date.available2009-08-27T16:14:04Z-
dc.date.issued2003-01-
dc.identifier.citationA new missense mutation in the growth hormone-releasing hormone receptor gene in familial isolated GH deficiency. 2003, 148 (1):25-30 Eur. J. Endocrinol.en
dc.identifier.issn0804-4643-
dc.identifier.pmid12534354-
dc.identifier.urihttp://hdl.handle.net/10541/79013-
dc.description.abstractOBJECTIVE: Mutations in the GH-releasing hormone (GHRH) receptor (GHRHR) gene (GHRHR) cause autosomal recessive familial isolated GH deficiency (IGHD). We searched for GHRHR mutations in two siblings with IGHD type IB and a history of parental consanguinity. DESIGN: We analyzed peripheral genomic DNA of an index patient. After identifying a novel mutation in the GHRHR, we performed functional studies in order to confirm that the mutation causes receptor malfunction. METHODS: The entire GHRHR was analyzed in the index case by denaturing gradient gel electrophoresis. Abnormally migrating bands were isolated and sequenced. The mutated area was then sequenced in all family members whose DNA was available. The newly found mutation was inserted into a GHRHR cDNA. Wild-type and mutant cDNAs were expressed into CHO cells and the cyclic AMP (cAMP) response to GHRH was measured. In order to determine whether the mutant receptor was properly expressed on the cell membrane surface, CHO cells were transfected with wild-type or mutant GHRHR cDNA containing a FLAG epitope tag in the extracellular N-terminus. RESULTS: Both patients were homozygous for a new missense mutation in codon 176, corresponding to the second transmembrane domain of the receptor protein that replaces alanine with valine (A176V). The mother and three unaffected siblings were heterozygous for the mutation; DNA from the father was not available. Cells expressing the A176V receptor had a significantly reduced cAMP response to GHRH, despite appropriate expression on the cell surface. CONCLUSIONS: We describe two siblings with IGHD due to a new mutation in the GHRHR that disrupts GHRH signaling and leads to GHRH resistance.en
dc.language.isoenen
dc.subject.meshChild-
dc.subject.meshDNA Mutational Analysis-
dc.subject.meshFamily Health-
dc.subject.meshFemale-
dc.subject.meshGene Expression-
dc.subject.meshHuman Growth Hormone-
dc.subject.meshHumans-
dc.subject.meshMale-
dc.subject.meshMutation, Missense-
dc.subject.meshPedigree-
dc.subject.meshReceptors, Neuropeptide-
dc.subject.meshReceptors, Pituitary Hormone-Regulating Hormone-
dc.subject.meshSiblings-
dc.titleA new missense mutation in the growth hormone-releasing hormone receptor gene in familial isolated GH deficiency.en
dc.typeArticleen
dc.contributor.departmentDivisions of Pediatric Endocrinology and Endocrinology and The Ilyssa Center for Molecular and Cellular Endocrinology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.en
dc.identifier.journalEuropean Journal of Endocrinologyen

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