title: Systematic screening for PRKAR1A gene rearrangement in Carney complex: identification and functional characterization of a new in-frame deletion.
abstract: BACKGROUND
  Point mutations of the PRKAR1A gene are a genetic cause of Carney complex (CNC) and primary pigmented nodular adrenocortical disease (PPNAD), but in 30% of the patients no mutation is detected.
  
  
  OBJECTIVE
  Set up a routine-based technique for systematic detection of large deletions or duplications of this gene and functionally characterize these mutations.
  
  
  METHODS
  Multiplex ligation-dependent probe amplification (MLPA) of the 12 exons of the PRKAR1A gene was validated and used to detect large rearrangements in 13 typical CNC and 39 confirmed or putative PPNAD without any mutations of the gene. An in-frame deletion was characterized by western blot and bioluminescence resonant energy transfer technique for its interaction with the catalytic subunit.
  
  
  RESULTS
  MLPA allowed identification of exons 3-6 deletion in three patients of a family with typical CNC. The truncated protein is expressed, but rapidly degraded, and does not interact with the protein kinase A catalytic subunit.
  
  
  CONCLUSIONS
  MLPA is a powerful technique that may be used following the lack of mutations detected by direct sequencing in patients with bona fide CNC or PPNAD. We report here one such new deletion, as an example. However, these gene defects are not a frequent cause of CNC or PPNAD.
authors.affiliation: Département de Biologie Hormonale, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 27 Rue du Faubourg Saint Jacques, 75014 Paris, France.|INSERM U970, Université Paris Descartes, PARCC, 56 Rue Leblanc, 75015 Paris, France.
authors.forename: M Guillaud
authors.initials: MG
authors.lastname: Bataille
authors.affiliation: Département de Biologie Hormonale, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 27 Rue du Faubourg Saint Jacques, 75014 Paris, France.|INSERM U970, Université Paris Descartes, PARCC, 56 Rue Leblanc, 75015 Paris, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Rhayem
authors.affiliation: Serviço de Genetica Medica, Departamento Pediatrico, Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal.|Clinical and Molecular Genetics Unit, UCL Institute of Child Health, London, UK.
authors.forename: S B
authors.initials: SB
authors.lastname: Sousa
authors.affiliation: Service d'Endocrinologie, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 75014 Paris, France.
authors.forename: R
authors.initials: R
authors.lastname: Libé
authors.affiliation: Département de Biologie Hormonale, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 27 Rue du Faubourg Saint Jacques, 75014 Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Dambrun
authors.affiliation: INSERM U970, Université Paris Descartes, PARCC, 56 Rue Leblanc, 75015 Paris, France.
authors.forename: C
authors.initials: C
authors.lastname: Chevalier
authors.affiliation: Département de Biologie Hormonale, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 27 Rue du Faubourg Saint Jacques, 75014 Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Nigou
authors.affiliation: INSERM U970, Université Paris Descartes, PARCC, 56 Rue Leblanc, 75015 Paris, France.
authors.forename: C
authors.initials: C
authors.lastname: Auzan
authors.affiliation: Département de Biologie Hormonale, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 27 Rue du Faubourg Saint Jacques, 75014 Paris, France.
authors.forename: M O
authors.initials: MO
authors.lastname: North
authors.affiliation: Serviço de Genetica Medica, Departamento Pediatrico, Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal.
authors.forename: J
authors.initials: J
authors.lastname: Sa
authors.affiliation: Serviço de Endocrinologia.
authors.forename: L
authors.initials: L
authors.lastname: Gomes
authors.affiliation: Section on Endocrinology and Genetics, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, Maryland 20892, USA.
authors.forename: P
authors.initials: P
authors.lastname: Salpea
authors.affiliation: Section on Endocrinology and Genetics, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, Maryland 20892, USA.
authors.forename: A
authors.initials: A
authors.lastname: Horvath
authors.affiliation: Section on Endocrinology and Genetics, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, Maryland 20892, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Stratakis
authors.affiliation: Département de Biologie Hormonale, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 27 Rue du Faubourg Saint Jacques, 75014 Paris, France.
authors.forename: N
authors.initials: N
authors.lastname: Hamzaoui
authors.affiliation: Service d'Endocrinologie, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 75014 Paris, France.|INSERM U1060, CNRS, Institut Cochin, Université Paris Descartes, Paris, France.
authors.forename: J
authors.initials: J
authors.lastname: Bertherat
authors.affiliation: Département de Biologie Hormonale, Hôpital Cochin, Assistance Publique - Hôpitaux de Paris, 27 Rue du Faubourg Saint Jacques, 75014 Paris, France.|INSERM U970, Université Paris Descartes, PARCC, 56 Rue Leblanc, 75015 Paris, France.
authors.forename: E
authors.initials: E
authors.lastname: Clauser
mesh_terms: D000293:Adolescent; D000303:Adrenal Cortex Diseases / Q000235:genetics / Q000378:metabolism; D000328:Adult; D056733:Carney Complex / Q000235:genetics* / Q000378:metabolism; D020134:Catalytic Domain; D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D005091:Exons; D005192:Family Health; D005260:Female; D017353:Gene Deletion; D015321:Gene Rearrangement; D056726:Genetic Association Studies; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D055550:Protein Stability; D055815:Young Adult
chemical_list: D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit; C517834:PRKAR1A protein, human; D010446:Peptide Fragments; D017868:Cyclic AMP-Dependent Protein Kinases