CHD8 interacts with CHD7, a protein which is mutated in CHARGE syndrome.
CHARGE syndrome is an autosomal dominant disorder caused in about two-third of cases by mutations in the CHD7 gene. For other genetic diseases e.g. hereditary spastic paraplegia, it was shown that interacting partners are involved in the underlying cause of the disease. These data encouraged us to search for CHD7 binding partners by a yeast two-hybrid library screen and CHD8 was identified as an interacting partner. The result was confirmed by a direct yeast two-hybrid analysis, co-immunoprecipitation studies and by a bimolecular fluorescence complementation assay. To investigate the function of CHD7 missense mutations in the CHD7-CHD8 interacting area on the binding capacity of both proteins, we included three known missense mutations (p.His2096Arg, p.Val2102Ile and p.Gly2108Arg) and one newly identified missense mutation (p.Trp2091Arg) in the CHD7 gene and performed both direct yeast two-hybrid and co-immunoprecipitation studies. In the direct yeast two-hybrid system, the CHD7-CHD8 interaction was disrupted by the missense mutations p.Trp2091Arg, p.His2096Arg and p.Gly2108Arg, whereas in the co-immunoprecipitation studies disruption of the CHD7-CHD8 interaction by the mutations could not be observed. The results lead to the hypothesis that CHD7 and CHD8 proteins are interacting directly and indirectly via additional linker proteins. Disruption of the direct CHD7-CHD8 interaction might change the conformation of a putative large CHD7-CHD8 complex and could be a disease mechanism in CHARGE syndrome.
Batsukh
Tserendulam
T
Institute of Human Genetics, University of Göttingen, 37073 Göttingen, Germany.
Pieper
Lasse
L
Koszucka
Anna M
AM
von Velsen
Nina
N
Hoyer-Fender
Sigrid
S
Elbracht
Miriam
M
Bergman
Jorieke E H
JE
Hoefsloot
Lies H
LH
Pauli
Silke
S
D000015:Abnormalities, Multiple / Q000235:genetics* / Q000378:metabolism; D002754:Choanal Atresia / Q000150:complications / Q000235:genetics / Q000378:metabolism; D003103:Coloboma / Q000150:complications / Q000235:genetics / Q000378:metabolism; D004265:DNA Helicases / Q000235:genetics* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D003638:Deafness / Q000150:complications / Q000151:congenital / Q000235:genetics / Q000378:metabolism; D002658:Developmental Disabilities / Q000150:complications / Q000235:genetics / Q000378:metabolism; D004423:Ear / Q000002:abnormalities; D006367:HeLa Cells; D006330:Heart Defects, Congenital / Q000150:complications / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009154:Mutation / Q000502:physiology; D011485:Protein Binding / Q000235:genetics; D054730:Protein Interaction Domains and Motifs / Q000235:genetics / Q000502:physiology; D050035:Sexual Infantilism / Q000150:complications / Q000235:genetics / Q000378:metabolism; D013577:Syndrome; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques
C523331:CHD8 protein, human; D004268:DNA-Binding Proteins; D014157:Transcription Factors; D004265:DNA Helicases; C491434:CHD7 protein, human