The transition from spermatogonia to spermatocytes and the initiation of meiosis are key steps in spermatogenesis and are precisely regulated by a plethora of proteins. However, the underlying molecular mechanism remains largely unknown. Here, we report that Src homology domain tyrosine phosphatase 2 (Shp2; encoded by the protein tyrosine phosphatase, nonreceptor type 11 [Ptpn11] gene) is abundant in spermatogonia but markedly decreases in meiotic spermatocytes. Conditional knockout of Shp2 in spermatogonia in mice using stimulated by retinoic acid gene 8 (Stra8)-cre enhanced spermatogonial differentiation and disturbed the meiotic process. Depletion of Shp2 in spermatogonia caused many meiotic spermatocytes to die; moreover, the surviving spermatocytes reached the leptotene stage early at postnatal day 9 (PN9) and the pachytene stage at PN11-13. In preleptotene spermatocytes, Shp2 deletion disrupted the expression of meiotic genes, such as disrupted meiotic cDNA 1 (Dmc1), DNA repair recombinase rad51 (Rad51), and structural maintenance of chromosome 3 (Smc3), and these deficiencies interrupted spermatocyte meiosis. In GC-1 cells cultured in vitro, Shp2 knockdown suppressed the retinoic acid (RA)-induced phosphorylation of extracellular-regulated protein kinase (Erk) and protein kinase B (Akt/PKB) and the expression of target genes such as synaptonemal complex protein 3 (Sycp3) and Dmc1. Together, these data suggest that Shp2 plays a crucial role in spermatogenesis by governing the transition from spermatogonia to spermatocytes and by mediating meiotic progression through regulating gene transcription, thus providing a potential treatment target for male infertility.
Animals ; Cell Cycle Proteins/genetics* ; Cell Line ; Cell Survival ; Chondroitin Sulfate Proteoglycans/genetics* ; Chromosomal Proteins, Non-Histone/genetics* ; Gene Expression Regulation ; Gene Knockdown Techniques ; Infertility, Male ; Male ; Meiosis/genetics* ; Mice ; Mice, Knockout ; Mice, Transgenic ; Phosphate-Binding Proteins/genetics* ; Protein Tyrosine Phosphatase, Non-Receptor Type 11/genetics* ; Rad51 Recombinase/genetics* ; Real-Time Polymerase Chain Reaction ; Spermatocytes/metabolism* ; Spermatogenesis/genetics* ; Spermatogonia/metabolism*

Animals ; Chromosomal Proteins, Non-Histone ; chemistry ; genetics ; metabolism ; Humans ; Models, Molecular ; Mutation ; Transcription Factors ; chemistry ; genetics ; metabolism

Chromosomal Proteins, Non-Histone ; chemistry ; genetics ; metabolism ; HeLa Cells ; Histones ; chemistry ; genetics ; metabolism ; Humans ; Protein Domains

MRG proteins are conserved during evolution in fungi, flies, mammals and plants, and they can exhibit diversified functions. The animal MRGs were found to form various complexes to activate gene expression. Plant MRG1/2 and MRG702 were reported to be involved in the regulation of flowering time via binding to H3K36me3-marked flowering genes. Herein, we determined the crystal structure of MRG701 chromodomain (MRG701). MRG701 forms a novel dimerization fold both in crystal and in solution. Moreover, we found that the dimerization of MRG chromodomains is conserved in green plants. Our findings may provide new insights into the mechanism of MRGs in regulation of gene expression in green plants.
Amino Acid Sequence ; Arabidopsis ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Binding Sites ; Chromosomal Proteins, Non-Histone ; chemistry ; genetics ; metabolism ; Cloning, Molecular ; Crystallography, X-Ray ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Histones ; chemistry ; genetics ; metabolism ; Models, Molecular ; Oryza ; genetics ; metabolism ; Peptides ; chemistry ; genetics ; metabolism ; Protein Binding ; Protein Interaction Domains and Motifs ; Protein Isoforms ; chemistry ; genetics ; metabolism ; Protein Multimerization ; Protein Structure, Secondary ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Sequence Homology, Amino Acid ; Viridiplantae ; genetics ; metabolism

OBJECTIVETo study the immunophenotype and molecular genetics of epithelioid sarcoma (ES), INI1 expression and its role in differential diagnosis.

METHODSTwenty cases of ES were retrieved from the archival files and selected for immunohistochemical study, DNA sequencing and fluorescence in-situ hybridization. The clinical and pathologic features were also reviewed.

RESULTSThe age of patients ranged from 16 to 75 years (mean = 40.2 years). The median age of patients in classic ES and proximal-type was 37.9 years and 42.0 years, respectively. The male-to-female ratio was 1.2: 1.0. Classic ES mostly occurred in the extremities while proximal-type ES often affected the perineum and external genitalia and trunk. Histologically, granuloma-like structures, consisting of aggregates of epithelioid and spindly tumor cells with central necrosis, were observed in classic ES. The epithelioid tumor cells contained abundant eosinophilic cytoplasm, merged with spindly cells at the periphery and admixed with collagen fibers. In proximal-type ES, the tumor cells showed prominent epithelioid and/or rhabdoid features, had marked cytologic atypia and grew in multinodular or diffuse patterns. In 2 cases of proximal-type ES studied, the "rhabdoid" tumor cells demonstrated a diffuse sheet-like growth pattern, mimicking malignant rhabdoid tumor. Immunohistochemical study showed that vimentin was positive in all cases. Pan-cytokeratin, CK8, CK7, epithelial membrane antigen and CD34 were expressed in 16, 15, 1, 18 and 13 cases, respectively. The staining for S-100 protein was focal and weak in 5 cases. None of the cases studied expressed CD31 and HMB45. Loss of INI1 was demonstrated in 10 of the 13 classic ES cases and 5 of the 7 proximal-type ES cases. The mutation of INI1 gene was detected in 1 of the 6 cases. Deletion of INI1 gene including heterozygous deletion, homozygous deletion and haploid was observed in 8 of the 11 cases.

CONCLUSIONSOwing to the histologic heterogeneity, pitfalls in diagnosis of ES sometimes are encountered. INI1 is lost in most cases of ES. Immunohistochemical study, including staining for INI1, provides useful clues in pathologic diagnosis. Instead of INI1 mutation, inactivation of INI1 gene related deletion is not uncommon.

Adolescent ; Adult ; Aged ; Antigens, CD34 ; metabolism ; Chromosomal Proteins, Non-Histone ; genetics ; DNA-Binding Proteins ; genetics ; Female ; Gene Deletion ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Keratins ; metabolism ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Mutation ; Rhabdoid Tumor ; genetics ; SMARCB1 Protein ; Sarcoma ; genetics ; Transcription Factors ; genetics ; Young Adult

Genetic information stored in DNA is accurately copied and transferred to subsequent generations through DNA replication. This process is accomplished through the concerted actions of highly conserved DNA replication components. Epigenetic information stored in the form of histone modifications and DNA methylation, constitutes a second layer of regulatory information important for many cellular processes, such as gene expression regulation, chromatin organization, and genome stability. During DNA replication, epigenetic information must also be faithfully transmitted to subsequent generations. How this monumental task is achieved remains poorly understood. In this review, we will discuss recent advances on the role of DNA replication components in the inheritance of epigenetic marks, with a particular focus on epigenetic regulation in fission yeast. Based on these findings, we propose that specific DNA replication components function as key regulators in the replication of epigenetic information across the genome.
Cdc20 Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Centromere ; metabolism ; Chromatin ; metabolism ; Chromosomal Proteins, Non-Histone ; metabolism ; DNA Replication ; DNA, Fungal ; metabolism ; Epigenesis, Genetic ; Histones ; metabolism ; Schizosaccharomyces ; genetics ; metabolism ; Schizosaccharomyces pombe Proteins ; antagonists & inhibitors ; genetics ; metabolism

OBJECTIVETo explore the role of centromere protein H (CENP-H) in the proliferation of human gastric cancer cells.

METHODSRT-PCR and Western blot analysis were employed to examine the mRNA and protein expressions of CENP-H in 7 human gastric cancer cell lines and immortalized human gastric epithelial cells (GES-1). The cells were infected with the retrovirus vectors pMSCV-CENP-H or CENP-H-RNAi to establish stable cell lines with high CENP-H expression or CENP-H expression interference. MTT assay and colony formation assay were used to examine the changes in the cell proliferation after the infection.

RESULTSCENP-H was over-expressed in gastric cancer cell lines AGS, BGC823, SGC-7901, MKN45, HGC27, MGC-803 and MKN28 at both mRNA and protein levels. The established AGS/CENP-H cell line with increased CENP-H expression showed enhanced proliferative activity, while the cell line MGC-803/CENP-H-RNAi with CENP-H expression interference showed an obviously lowered proliferation ability.

CONCLUSIONCENP-H promotes the proliferation of human gastric cancer cells, suggesting its important role in the occurrence and development of gastric cancer.

Cell Line, Tumor ; Cell Proliferation ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Humans ; RNA, Messenger ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Stomach Neoplasms ; metabolism ; pathology

BACKGROUNDIntegrase interactor 1 (INI1), which encodes a component of the ATP-dependent chromatin remodeling hSWI-SNF complex, has been identified as a tumor suppressor in many tumors. Nonetheless, the role of INI1 in gastric tumor progression is not known exactly. The aim of this research was to investigate the effect of INI1 in the carcinogenesis and progression of gastric cancer.

METHODSGastric tumor tissues with different differentiation levels from clinical gastric carcinoma samples and adjacent control normal tissues were taken. Expression levels of INI1 were detected by quantitative reverse transcriptation-polymerase chain reaction (RT-PCR) and Western blotting. Gastric cancer cell line SGC7901 was transfected with INI1 eukaryotic expressing vector INI1-GFP. Cell proliferation activities were assessed by MTT; cell count and cell cycle were detected by flow cytometry (FCM); cell apoptosis were measured by TUNEL and FCM; cell migration and invasiveness were evaluated by wound healing and transwell assays. Expression levels of INI1 and proliferation-related genes including p16, p21, cyclin D1 and cyclin A, apoptosis genes p53, B-cell non-Hodgkin lymphoma-2 (Bcl-2), Bcl-2-associated x protein (Bax) and caspase-3, and invasion-related genes including intercellular adhesion molecule 1 (ICAM1), matrix metalloproteinase 2 (MMP2), MMP9 and tissue inhibitor of matrix metalloproteinase 1 (TIMP1), were detected by quantitative RT-PCR and Western blotting.

RESULTSINI1 expression levels were lower in gastric carcinoma compared with adjacent control normal tissues. Overexpression of INI1 in SGC7901 cells inhibited its proliferation and invasiveness, but increased anoikis and G(0)/G(1) cell number. INI1-GFP transfection upregulated expression of INI1 and proliferation related genes p16 and p21, apoptosis genes p53 and Bax, and invasion-related genes TIMP1; cyclin D1, cyclin A, Bcl2, ICAM1, MMP2 and MMP9 were downregulated, and there was no significant change in caspase 3 levels.

CONCLUSIONINI1 plays a key role in gastric carcinogenesis by affecting proliferation, apoptosis and invasion.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Cycle ; genetics ; physiology ; Cell Line, Tumor ; Cell Proliferation ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Humans ; Real-Time Polymerase Chain Reaction ; SMARCB1 Protein ; Stomach Neoplasms ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

Astrocytoma ; genetics ; metabolism ; pathology ; Brain Neoplasms ; classification ; genetics ; metabolism ; pathology ; Child ; Child, Preschool ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Chromosome Deletion ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Ependymoma ; genetics ; metabolism ; pathology ; Hedgehog Proteins ; genetics ; metabolism ; Humans ; Medulloblastoma ; classification ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins B-raf ; genetics ; metabolism ; Rhabdoid Tumor ; genetics ; metabolism ; pathology ; SMARCB1 Protein ; Signal Transduction ; Transcription Factors ; genetics ; metabolism ; Wnt Proteins ; metabolism ; beta Catenin ; genetics ; metabolism

OBJECTIVETo determine the correlation between the expression of CARMA1 mRNA and MUM1 protein, as well as its effects on clinicopathological features and prognosis of diffuse large B cell lymphoma (DLBCL).

METHODSThe immunophenotype (CD20, CD79a, CD10, MUM1, Bcl6) and proliferation index of DLBCL cells were examined by immunohistochemistry (IHC). CARMA1 mRNA was detected by RT-PCR.

RESULTSCARMA1 mRNA was detected in 76 of 89 (85.40%) cases with DLBCL. The level of CARMA1 mRNA was higher in MUM1-postive group than in MUM1-negative group. No correlation was found in the expression intensity between the two molecules (P = 0.084). Ki67 positive rate was higher in MUM1(+) cases than in MUM1(-) ones (P = 0.030). There was no difference between MUM1(+) and MUM1(-) cases in sex, median age, staging, primary site and other clinicopathological features. In 58 CARMA1 mRNA positive cases, low expression cases showed more in earlier stage and more males. No difference in survival status was identified between cases with and without MUM1 expression, over- and low-expression of CARMA1 mRNA, as well as over- and low-expression of CARMA1 mRNA among 58 cases with MUM1 expression.

CONCLUSIONThe expression of CARMA1 mRNA is likely associated with the expression of MUM1 and shows male predominance in DLBCL. The expression of CARMA1 may be involved with pathogenesis and progression of ABC-like DLBCL. The two molecules correlated somewhat with some clinicopathological features, but not with survival of DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; CARD Signaling Adaptor Proteins ; genetics ; metabolism ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Female ; Guanylate Cyclase ; genetics ; metabolism ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; pathology ; Neoplasm Staging ; Young Adult