mPem, a homeobox gene, is expressed in a time and stage specific manner during murine ontogeny. Pem transcripts are abundant in 7- and 8-day mouse embryos, but decrease precipitously thereafter. On Day 9 they become abundant in placenta and yolk sac, persisting there until parturition. Although Pem transcripts are not detectable in most of adult tissues, they are present in reproductive system such as testis, epididymis and ovary. This indicates a important role for Pem during embryogenesis and reproductive development. To study the function of mPem protein, we used a GAL4 based yeast two-hybrid assay to screen a 7-day mouse embryo library with full-length of mPem. 3 proteins were found interacting with mPem protein. One of theses is Mdfic. We confirmed the interaction between mPem and Mdfic in yeast and in vitro. Mdfic, MyoD family inhibitor domain containing, encodes the myoD family inhibitor domain (I-mfa domain). The interaction between mPem and Mdfic suggested they maybe form the transcriptional regulator complex to regulate embryo differentiation.
Animals ; Embryo, Mammalian ; Embryonic Development ; Female ; Genes, Homeobox ; genetics ; Homeodomain Proteins ; chemistry ; metabolism ; Mice ; Pregnancy ; Protein Binding ; Transcription Factors ; chemistry ; metabolism ; Two-Hybrid System Techniques

OBJECTIVETo screen the variations of TG interacting factor(TGIF) gene in encoding sequence in Chinese high myopia patients and normal controls and to analyze the SNPs of TGIF gene encoding sequence in Chinese population.

METHODSGenomic DNA was collected from 204 probands with high myopia and 112 unrelated persons without high myopia. The coding sequences of TGIF gene in 316 subjects were analyzed by using exon-by-exon PCR heteroduplex-SSCP analysis and sequencing.

RESULTSThere were 3 types of SNP and one single nucleotide mutation in the coding sequence of TGIF gene: IVS-2 nt350 G --> T(36/204), codon140 CCA --> CCG; Pro140Pro codon163 CCG --> CTG;Pro163Leu and codon126 GTG --> GCG; Val126Ala(1/204). The SNPs of codon140 CCA --> CCG and codon163 CCG --> CTG were composed of 3 alleles and 5 genotypes in Chinese population which abide by Hardy-Weinberg law.

CONCLUSIONThere was no evidence to prove that mutations in the TGIF gene are responsible for the high myopia in Chinese. Three SNPs of coding sequence TGIF gene in Chinese population abide by Hardy-Weinberg law.

China ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Gene Frequency ; Homeodomain Proteins ; genetics ; Humans ; Mutation ; Myopia ; genetics ; pathology ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational ; Repressor Proteins ; genetics

Axenfeld-Rieger syndrome (ARS) is characterized by anomalies of the anterior segment of the eye and systemic abnormalities. Mutations in the FOXC1 and PITX2 genes are underlying causes of ARS, but there has been few reports on genetically confirmed ARS in Korea. We identified a novel PITX2 mutation (c.300_301delinsT) in 2 Korean patients from a family with ARS. We expand the spectrum of PITX2 mutations and, to the best of our knowledge, this is the first confirmed family of PITX2-related ARS in Korea.
Adult ; Anterior Eye Segment/*abnormalities/pathology ; Base Sequence ; Child, Preschool ; Eye Abnormalities/*genetics/pathology ; Female ; Heterozygote ; Homeodomain Proteins/chemistry/*genetics ; Humans ; Mutation ; Pedigree ; Republic of Korea ; Transcription Factors/chemistry/*genetics

Nail-patella syndrome (NPS) is an autosomal dominant disease that typically involves the nails, knees, elbows and the presence of iliac horns. In addition, some patients develop glomerulopathy or adult-onset glaucoma. NPS is caused by lossof- function mutations in the LMX1B gene. In this study, phenotype-genotype correlation was analyzed in 9 unrelated Korean children with NPS and their affected family members. The probands included 5 boy and 4 girls who were confirmed to have NPS, as well as 6 of their affected parents. All of the patients (100%) had dysplastic nails, while 13 patients (86.7%) had patellar anomalies, 8 (53.3%) had iliac horns, 6 (40.0%) had elbow contracture, and 4 (26.7%) had nephropathy including one patient who developed end-stage renal disease at age 4.2. The genetic study revealed 8 different LMX1B mutations (5 missense mutations, 1 frame-shifting deletion and 2 abnormal splicing mutations), 6 of which were novel. Genotype-phenotype correlation was not identified, but inter- and intrafamilial phenotypic variability was observed. Overall, these findings are similar to the results of previously conducted studies, and the mechanism underlying the phenotypic variations and predisposing factors of the development and progression of nephropathy in NPS patients are still unknown.
Adolescent ; Child ; Child, Preschool ; DNA Primers/chemistry ; Female ; Genotype ; Homeodomain Proteins/*genetics ; Humans ; Infant ; Kidney Failure, Chronic/genetics ; Korea ; Male ; Mutation ; Nail-Patella Syndrome/diagnosis/*genetics/physiopathology ; Phenotype ; Transcription Factors/*genetics

BACKGROUNDNeural tube defects are the most common human birth defects. The causes are multifactorial with complex genetic and environmental factors, although the exact genetic causes are unknown. This research was conducted to study the frequency of Msx2 gene polymorphisms in 59 women with a history of pregnancy with a neural tube defect and in 73 healthy controls. We aimed to determine the effect of this genetic polymorphism on the incidence of neural tube defects in the Han Chinese population.

METHODSWe studied 59 mothers with at least one previous child with a neural tube defect (the case group) and 73 case-control subjects during the same period, from Shanxi Province, China. We analyzed the genotypic distributions and allele frequencies of Msx2 C386T polymorphisms in DNA samples from the case and control groups. A three-dimensional protein model was predicted using Swiss-Pdb Viewer software version 4.0. Disease association was analyzed using chi-square tests.

RESULTSSignificant differences were observed in the genotypes and allele frequencies of the Msx2 C386T allele between the case and control groups (CT: 32% vs. 15%, P = 0.0073 and TT 15% vs. 4%, P = 0.013, respectively). Logistic regression analysis showed that the C386T mutation is a potential risk factor for neural tube defects (P < 0.05; OR: 3.466; 95%CI: 1.831 - 6.560). Three-dimensional structure prediction revealed that the Msx2 C386T mutation results in a threonine substitution for methionine at position 129 of exon 2, which might lead to structural mutations or dysfunctions in the protein encoded by Msx2.

CONCLUSIONMaternal Msx2 C386T gene polymorphisms were associated with fetal neural tube defects in Han Chinese women in Shanxi Province.

Adult ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Homeodomain Proteins ; chemistry ; genetics ; metabolism ; Humans ; Neural Tube Defects ; epidemiology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; genetics ; Pregnancy ; Protein Structure, Secondary ; Young Adult

OBJECTIVETo study the role of a BTB/POZ domain protein in the expression of hsp90alpha gene.

METHODSThe eukaryotic expression plasmids of sense- and antisense-GAGA related protein (GRP) or empty vector were transfected into Jurkat cells with pREP4 episomal vector plasmids carrying the hsp90alpha promoter sequence from -1756 to +37 and control plasmids pMCAT. Total RNA was extracted. The relative promoter activity of hsp90alpha-CAT reporter gene was determined by competitive RT-PCR assay.

RESULTSGRP markly increased the relative promoter activity of hsp90alpha-CAT reporter gene during heat shock.

CONCLUSIONGRP may promote the expression of hsp90alpha gene by participating in chromatin remolding.

Amino Acid Motifs ; Animals ; Checkpoint Kinase 1 ; Cloning, Molecular ; DNA ; metabolism ; DNA, Complementary ; metabolism ; DNA-Binding Proteins ; metabolism ; Drosophila ; genetics ; Drosophila Proteins ; genetics ; Gene Expression Regulation ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Heat-Shock Response ; genetics ; Homeodomain Proteins ; chemistry ; genetics ; metabolism ; Humans ; Repressor Proteins ; genetics ; Transcription Factors ; genetics ; Two-Hybrid System Techniques

Nanog, a homeodomain (HD) transcription factor, plays a critical role in the maintenance of embryonic stem (ES) cell self-renewal. Here, we report the identification of an alternatively-spliced variant of nanog. This variant lacked a stretch of amino acids (residues 168-183) located between the HD and tryptophan-repeat (WR) of the previously-reported full length sequence, suggesting that the deleted sequence functions as a linker and possibly affects the flexibility of the C-terminal transactivation domain relative to the DNA binding domain. Expression of mRNA encoding the splice variant, designated as nanog-delta 48, was much lower than that of the full length version in human ES cells. The ratio of nanog-delta 48 transcript to full length transcript increased, however, in multipotent adult progenitor cells. EMSA analysis revealed that both forms of Nanog were able to bind a nanog binding sequence with roughly the same affinity. A reporter plasmid assay also showed that both variants of nanog modestly repressed transactivation of gata-4, whose expression is proposed to be inhibited by nanog, with comparable potency. We conclude that, despite the difference in primary structure and expression pattern in various stem cells, the alternatively-spliced variant of Nanog has similar activity to that of the full length version.
Alternative Splicing/*genetics ; Amino Acid Sequence ; Base Sequence ; Cell Nucleus ; Cells, Cultured ; DNA-Binding Proteins/chemistry/*genetics/metabolism ; Exons/*genetics ; GATA4 Transcription Factor/metabolism ; Gene Expression Profiling ; Genes, Reporter ; Homeodomain Proteins/chemistry/*genetics/metabolism ; Humans ; Introns/genetics ; Molecular Sequence Data ; Promoter Regions (Genetics) ; RNA, Messenger/genetics/metabolism ; Research Support, Non-U.S. Gov't ; Trans-Activation (Genetics) ; Transfection

To investigate the effect and possible mechanism of echinacoside-containing serum on the osteogenic differentiation in rat bone marrow mesenchymal stem cells. Rat bone marrow mesenchymal stem cells were cultivated by the whole bone marrow adherence method. The 3rd generation of cells were divided into 3 groups: the blank control group, the classic osteogenic-induced group and the 10% echinacoside-containing serum group. The expression of alkaline phosphatase and osteocalcin were detected by ELISA. The ex- pression of ZHX, protein was detected by Western blot technique. RT-PCR technique was used to detect the expression of ZHX₃mRNA. According to the result, the expressions of the alkaline phosphatase and osteocalcin in the classic osteogenic-induced group and the 10% echinacoside-containing serum group were significantly higher than that of the blank control group (P <0. 01). And expressions of the alkaline phosphatase activity and osteocalcin in the 10% echinacoside-containing serum group were significantly higher than that in the classic osteogenic-induced group (P < 0.01). Meanwhile, the classic osteogenic-induced group and the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the black control group, with significant differences (P < 0.01); the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the classic osteogenic-induced group, with a significant difference (P < 0.01). In conclusion, 10% echinacoside-containing serum can promote the differentiation of the bone marrow mesenchymal stem cells cultured in vitro. Its mechanism may be correlated with the increase in the ZHX₃expression.
Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Glycosides ; blood ; pharmacology ; Homeodomain Proteins ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry ; Transcription Factors ; genetics ; metabolism

Animals ; Apoptosis ; Cell Cycle ; Cell Cycle Proteins ; genetics ; metabolism ; physiology ; DNA Repair ; Homeodomain Proteins ; genetics ; metabolism ; physiology ; Humans ; Inhibitor of Growth Protein 1 ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; physiology ; Neoplasm Metastasis ; Neoplasms ; metabolism ; pathology ; Neovascularization, Pathologic ; pathology ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Prognosis ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; physiology ; Signal Transduction ; Transcription Factors ; genetics ; metabolism ; physiology ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; chemistry ; genetics ; metabolism ; physiology