title: Novel IRF6 mutations in Japanese patients with Van der Woude syndrome: two missense mutations (R45Q and P396S) and a 17-kb deletion.
abstract: Three Japanese families with Van der Woude syndrome (VWS) were screened for mutations in the interferon regulatory factor 6 gene (IRF6) by sequencing its entire coding region. Two novel missense mutations, R45Q in exon 3 and P396S in exon 9, were identified in families 1 and 2, respectively. In family 3, no causative base change was found by the sequencing analysis, but a deletion involving exons 4-9 was suggested by multiplex PCR analysis. To confirm the deletion and to determine its 5'- and 3'-boundaries, we amplified a DNA fragment containing a heterozygous polymorphic site in exon 2 by using a 5'-upstream forward PCR primer and eight different reverse primers located 3'-downstream of exon 2. The amplified product was subjected to nested PCR to generate a DNA fragment containing the polymorphic site. When a reverse primer located within the deletion was used for the first PCR amplification, only the nondeletion allele was detected after the second PCR. Repeated analyses with eight different reverse primers allowed us to map the boundaries of the deletion, and subsequently a heterozygous 17,162-bp deletion involving exons 4-9 was identified. Since IRF6 mutations in a significant portion of VWS patients remain undetected by conventional sequencing analysis, it may be important to search for a large deletion in those patients. Our simple methods to identify deletions and to determine the boundaries of a deletion would facilitate the identification of such patients.
authors: Department of Medical Genetics, Tohoku University School of Medicine, Seiryomachi, Aoba-ku, Sendai 980-8574, Japan.|Department of Plastic and Reconstructive Surgery, Tohoku University School of Medicine, Sendai, Japan.
authors: Shuji
authors: S
authors: Kayano
authors: Department of Medical Genetics, Tohoku University School of Medicine, Seiryomachi, Aoba-ku, Sendai 980-8574, Japan. skure@mail.cc.tohoku.ac.jp.
authors: Shigeo
authors: S
authors: Kure
authors: Department of Medical Genetics, Tohoku University School of Medicine, Seiryomachi, Aoba-ku, Sendai 980-8574, Japan.
authors: Yoichi
authors: Y
authors: Suzuki
authors: Department of Medical Genetics, Tohoku University School of Medicine, Seiryomachi, Aoba-ku, Sendai 980-8574, Japan.|Department of Plastic and Reconstructive Surgery, Tohoku University School of Medicine, Sendai, Japan.
authors: Kiyoshi
authors: K
authors: Kanno
authors: Department of Medical Genetics, Tohoku University School of Medicine, Seiryomachi, Aoba-ku, Sendai 980-8574, Japan.
authors: Yoko
authors: Y
authors: Aoki
authors: Department of Pediatrics, The University of Iowa, Iowa City, Iowa, 52242, USA.
authors: Shinji
authors: S
authors: Kondo
authors: Department of Pediatrics, The University of Iowa, Iowa City, Iowa, 52242, USA.
authors: Brian C
authors: BC
authors: Schutte
authors: Department of Pediatrics, The University of Iowa, Iowa City, Iowa, 52242, USA.
authors: Jeffrey C
authors: JC
authors: Murray
authors: Department of Plastic and Reconstructive Surgery, Tohoku University School of Medicine, Sendai, Japan.
authors: Atsushi
authors: A
authors: Yamada
authors: Department of Medical Genetics, Tohoku University School of Medicine, Seiryomachi, Aoba-ku, Sendai 980-8574, Japan.
authors: Yoichi
authors: Y
authors: Matsubara
mesh_terms: D002971:Cleft Lip / Q000235:genetics; D002972:Cleft Palate / Q000235:genetics; D017931:DNA Primers / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics*; D005091:Exons; D005260:Female; D017353:Gene Deletion; D006579:Heterozygote; D006801:Humans; D050835:Interferon Regulatory Factors; D007564:Japan; D008297:Male; D008957:Models, Genetic; D009154:Mutation; D020125:Mutation, Missense; D010375:Pedigree; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA; D013577:Syndrome; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; C109803:IRF6 protein, human; D050835:Interferon Regulatory Factors; D014157:Transcription Factors