Korean J Orthod.  2008 Apr;38(2):133-143. 10.4041/kjod.2008.38.2.133.

Characteristics of MSX1 gene in Korean nonsyndromic cleft lip and palate individuals

• Lee HK
• Kim SS
• Son WS

• KMID: 1481500
• DOI: http://doi.org/10.4041/kjod.2008.38.2.133

Abstract

OBJECTIVE
This study was performed to identify the characteristics of the MSX1 gene (locus chromosome 4p16) in Korean nonsyndromic cleft lip and palate (CL/P), which is assumed to be a major candidate gene acting as a causal factor in nonsyndromic CL/P and missing teeth.
METHODS
The 36 individuals (23 males and 13 females) who had visited the department of orthodontics at from 1998 to 2002 and who had nonsyndromic CL/P were included in the study. Using a PCR-based assay, the MSX1 gene was amplified, sequenced, and searched for inferred protein products (Reference: Homo sapiens MSX1, accession number AF426432 and NP_002439). The common single nucleotide polymorphisms were observed.
RESULTS
In exon 1, nucleotide "A" of the 253 basepair (bp) region was substituted for "G", and in the 255 bp region, nucleotide "G" was inserted. In exon 2, nucleotide "C" of the 11 bp region was substituted for "A", and "T" or "G" was inserted into the 351 bp region whereas "T" or "A" was inserted into the 352 bp region. In protein analysis, "Thr85Ala" missense mutation was found. The "Thr85Ala" missense mutation in this study is different from those of studies using subjects of other races.
CONCLUSIONS
The results suggest that there is specific mutation of MSX1 in Korean and it plays an important role in Korean nonsyndromic CL/P. However, any distinct genetic polymorphisms between CL/P with missing teeth in the cleft region and CL/P without missing teeth could not be found.

Keyword

MSX1 gene; Korean nonsyndromic cleft lip and palate; Missing teeth

MeSH Terms

Cleft Lip
Continental Population Groups
Exons
Humans
Male
Mutation, Missense
Orthodontics
Palate
Polymorphism, Genetic
Polymorphism, Single Nucleotide
Tooth

Figure

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  Fig 1 Two percent agarose gel electrophoresis of MSX1 gene PCR product. M, molecular weight marker; lane 1, 2, 3, 4, nonsyndromic cleft lip and palate subjects; bp, basepair.

• 

  Fig 2 Three different genetic features were exhibited, and were divided into patterns I, II and III. The sequences of pattern I were similar to the reference sequences, and most of the subjects belonged to pattern I. However, the sequences of pattern II and pattern III were very different from the reference sequences, and they also differed from each other.

• 

  Fig 3 Neighbour joining tree of MSX1 gene from ClustalW multiple sequence alignment program. Most of the subjects except 9 subjects belonged to pattern I. "#" means patient number in raw data.

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  Fig 4 The common single nucleotide polymorphisms (SNPs) of pattern I. In exon 1, nucleotide "A" of the 253 basepair region was substituted for "G", and in the 255 basepair region, nucleotide "G" was inserted.

• 

  Fig 5 The common SNPs of exon 2. In exon 2, nucleotide "C" of the 11 basepair region was substituted for "A".

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  Fig 6 The common SNPs in exon 2 of pattern I in 351,352 basepair region. "T" or "G" was inserted into the 351 basepair region whereas "T" or "A" was inserted into the 352 basepair region.

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  Fig 7 Protein sequence comparison in exon 1 of the MSX1 gene. In 85 region, Threonine was substituted for Alanine in all subjects. Sequence changes of amino acid by this protein mutation in the 85 region were continued into 279 region and then reverted to the normal structure from the 280 region.

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  Fig 8 A, The protein sequences of patient number 32 were considerably different from those of other subjects in pattern I; B, nucleotide sequences of patient number 32: nucleotide "T" was inserted into the 57 bp region; C, the "Lys20Gln" missense mutation was found.

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