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Identification of a tspy co-expression network associated with DNA hypomethylation and tumor gene expression in somatic cancers

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Journal of Genetics and Genomics 2016年第10期43卷 577-585页

作者： Tatsuo Kido Yun-Fai Chris Lau Division of Cell and Developmental Genetics Department of Medicine Veterans Affairs Medical Center and Institute for Human Genetics University of California

Testis specific protein Y-encoded（tspy） is a Y-located proto-oncogene predominantly expressed in normal male germ cells and various types of germ cell tumor. Significantly, tspy is frequently expressed in somatic cancers including liver cancer but not in adjacent normal tissues, suggesting that ectopic tspy expression could be associated with oncogenesis in non-germ cell cancers. Various studies demonstrated that tspy expression promotes growth and proliferation in cancer cells; however, its relationship to other oncogenic events in tspy-positive cancers remains unknown. The present study seeks to correlate tspy expression with other molecular features in clinical cancer samples, by analyses of RNA-seq transcriptome and DNA methylation data in the Cancer Genome Atlas（TCGA） database. A total of 53 genes,including oncogenic lineage protein 28 homolog B（LIN28B） gene and RNA-binding motif protein Y-linked（RBMY） gene, are identified to be consistently co-expressed with tspy, and have been collectively designated as the tspy co-expression network（TCN）. TCN genes were simultaneously activated in subsets of liver hepatocellular carcinoma（30%） and lung adenocarcinoma（10%） regardless of pathological stage, but only minimally in other cancer types. Further analysis revealed that the DNA methylation level was globally lower in the TCN-active than TCN-silent cancers. The specific expression and methylation patterns of TCN genes suggest that they could be useful as biomarkers for the diagnosis,prognosis and clinical management of cancers, especially those for liver and lung cancers, associated with tspy co-expression network genes.

关键词： Co-expression network DNA methylation Gene expression signature Cancer subclassification Y chromosome genes tspy Cancer/testis antigens

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Expression of the human tspy gene in the brains of transgenic mice suggests a potential role of this Y chromosome gene in neural functions

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Journal of Genetics and Genomics 2011年第5期38卷 181-191页

作者： Tatsuo Kido Stephanie Schubert Jrg Schmidtke Yun-Fai Chris Lau Division of Cell and Developmental Genetics Department of MedicineVA Medical Centerand Institute for Human GeneticsUniversity of California Institute of Human Genetics Hannover Medical School

The testis specific protein Y-encoded （tspy） is a member of tspy/SET/NAP1 superfamily, encoded within the gonadoblastoma locus on the Y chromosome. tspy shares a highly conserved SET/NAP-domain responsible for protein--protein interaction among tspy/SET/NAP 1 proteins. Accumulating data, so far, support the role of tspyas the gonadoblastoma gene, involved in germ cell tumorigenesis. The X-chromosome homolog of tspy, TSPX is expressed in various tissues at both fetal and adult stages, including the brain, and is capable of interacting with the multi-domain adapter protein CASK, thereby influencing the synaptic and transcriptional functions and developmental regulation of CASK in the brain and other neural tissues. Similar to TSPX, we demonstrated that tspy could interact with CASK at its SET/NAP-domain in cultured cells. Transgenic mice harboring a human tspygene and flanking sequences showed specific expression of the human tspytransgene in both testis and brain. The neural expression pattern of the human tspygene overlapped with those of the endogenous mouse Cask and Tspx gene. Similarly with TSPX, tspy was co-localized with CASK in neuronal axon fibers in the brain, suggesting a potential role（s） of tspy in development and/or physiology of the nervous system.

关键词： tspy TSPX CASK Y chromosome Neuron Testis

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Battle of the sexes: contrasting roles of testis-specific protein Y-encoded (tspy) and TSPX in human oncogenesis

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Asian Journal of Andrology 2019年第3期21卷 260-269页

作者： Yun-Fai Chris Lau Yunmin Li Tatsuo Kido Division of Cell and Developmental Genetics Department of Medicine VA Medical Center and Institute for Human Genetics University of California San Francisco CA 94121 USA

The Y-located testis-specific protein Y-encoded (tspy) and its X-homologue TSPX originated from the same ancestral gene, but act as a proto-oncogene and a tumor suppressor gene, respectively. tspy has specialized in male-specific functions, while TSPX has assumed the functions of the ancestral gene. Both tspy and TSPX harbor a conserved SET/NAP domain, but are divergent at flanking structures. Specifically, TSPX contains a C-terminal acidic domain, absent in tspy. They possess contrasting properties, in which tspy and TSPX, respectively, accelerate and arrest cell proliferation, stimulate and inhibit cyclin B-CDK1 phosphorylation activities, have no effect and promote proteosomal degradation of the viral HBx oncoprotein, and exacerbate and repress androgen receptor (AR) and constitutively active AR variant, such as AR-V7, gene transactivation. The inhibitory domain has been mapped to the carboxyl acidic domain in TSPX, truncation of which results in an abbreviated TSPX exerting positive actions as tspy. Transposition of the acidic domain to the C-terminus of tspy results in an inhibitory protein as intact TSPX. Hence, genomic mutations/aberrant splicing events could generate TSPX proteins with truncated acidic domain and oncogenic properties as those for tspy. Further, tspy is upregulated by AR and AR-V7 in ligand-dependent and ligand-independent manners, respectively, suggesting the existence of a positive feedback loop between a Y-located proto-oncogene and male sex hormone/receptors, thereby amplifying the respective male oncogenic actions in human cancers and diseases. TSPX counteracts such positive feedback loop. Hence, tspy and TSPX are homologues on the sex chromosomes that function at the two extremes of the human oncogenic spectrum.

关键词： androgen receptor AR-V7 cell cycle regulation cyclin B-CDK1 oncogene sex chromosomes sex differences TSPX tspy tumor suppressor

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ZFY和tspy基因在荷斯坦奶牛精子中表达定位及其应用的研究

ZFY和TSPY基因在荷斯坦奶牛精子中表达定位及其应用的研究

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作者：刘方华中农业大学

学位级别：硕士

利用X/Y精子分离技术改变家畜后代性别比例,将有效地提高畜牧业的生产和经济效率。目前,根据X/Y染色体DNA含量差异和利用流式细胞仪分离技术分离X/Y精子,在奶牛业一定范围内得到应用并产生了一定的效益;但因其成本较高、精子在分离过程... 详细信息

利用X/Y精子分离技术改变家畜后代性别比例,将有效地提高畜牧业的生产和经济效率。目前,根据X/Y染色体DNA含量差异和利用流式细胞仪分离技术分离X/Y精子,在奶牛业一定范围内得到应用并产生了一定的效益;但因其成本较高、精子在分离过程中受到损伤、受胎率较低、以及国际知识产权限制等而不能被广泛应用,故需要进一步优化和提高该分离技术,或建立其它有效的分离X/Y精子的技术。如果能根据X/Y精子膜上的差异表达蛋白,利用抗体抗原免疫反应对精子进行分离,有望大批量生产性控精子,同时减少对精子的伤害并降低分离成本,具有较好的市场前景。本研究以牛Y染色体中tspy、ZFY两个基因为研究对象,为建立X/Y精子分离技术分离X/Y精子提供参考,主要开展了以下研究:(1)分别分析这两个蛋白在荷斯坦奶牛精子中的定位;(2)分析这两个基因在荷斯坦奶性控X/Y精子中的蛋白表达特征;(3)分别利用这两个蛋白抗体分选荷斯坦奶牛X/Y性控精子,以探讨用于分离荷斯坦奶牛X/Y精子的可能性,为建立X/Y精子分离技术提供参考。主要研究结果如下:***基因主要定位在荷斯坦奶牛精子的尾部,信号较弱;tspy蛋白定位于在荷斯坦奶牛精子的顶体部分和细胞核内。***和tspy基因在性控Y精子的总蛋白中的表达量均极显著高于性控X精子(Ptspy抗体在荷斯坦奶牛性控Y精子中的结合率(41.75%±4.05%)极显著(Ptspy抗体分别分选阳性结合的精子和阴性结合的精子,结果显示,利用ZFY抗体分选出的阳性精子提高到77.7%,利用tspy抗体分选出的阳性精子提高到82.2%。这两个蛋白可以作为Y精子标识蛋白。综上所述,tspy定位于荷斯坦奶牛Y精子的顶体和细胞核内;ZFY主要定位在荷斯坦奶牛Y精子的尾部。而且,这两个基因可以作为Y精子标识蛋白在X/Y精子分离中具有较好的潜在应用价值。

关键词：荷斯坦奶牛性别特异蛋白 tspy ZFY

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Roles of the Y chromosome genes in human cancers

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Asian Journal of Andrology 2015年第3期17卷 373-380页

作者： Tatsuo Kido Yun-Fai Chris Lau Division of Cell and Developmental Genetics Department of Medicine Veterans Affairs Medical Center Institute for Human Genetics University of California San Francisco California 94121 USA

Male and female differ genetically by their respective sex chromosome composition, that is, XY as male and XX as female. Although both X and Y chromosomes evolved from the same ancestor pair of autosomes, the Y chromosome harbors male-specific genes, which play pivotal roles in male sex determination, germ cell differentiation, and masculinization of various tissues. Deletions or translocation of the sex-determining gene, SRY, from the Y chromosome causes disorders of sex development （previously termed as an intersex condition） with dysgenic gonads. Failure of gonadal development results not only in infertility, but also in increased risks of germ cell tumor （GCT）, such as gonadoblastoma and various types of testicular GCT. Recent studies demonstrate that either loss of Y chromosome or ectopic expression of Y chromosome genes is closely associated with various male-biased diseases, including selected somatic cancers. These observations suggest that the Y-linked genes are involved in male health and diseases in more frequently than expected. Although only a small number of protein-coding genes are present in the male-specific region of Y chromosome, the impacts of Y chromosome genes on human diseases are still largely unknown, due to lack of in vivo models and differences between the Y chromosomes of human and rodents. In this review, we highlight the involvement of selected Y chromosome genes in cancer development in men.

关键词： germ cell tumors RBMY, Y-linked somatic cancers tspy Y chromosome

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Absolute copy number differences of Y chromosomal genes between crossbred (Bos taurus×Bos indicus) and Indicine bulls

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Journal of Animal Science and Biotechnology 2013年第4期4卷 257-263页

作者： Ayan Mukherjee Gulshan Dass Jagan Mohanarao G Moloya Gohain Biswajit Brahma Tirtha Kumar Datta Sachinandan De Animal Genomics Lab Animal Biotechnology Centre National Dairy Research Institute Department of Veterinary Biochemistry College of Veterinary Sciences & Animal Husbandry Central Agricultural University

Background： The Y chromosome in mammal is paternally inherited and harbors genes related to male fertility and spermatogenesis. The unique intra-chromosomal recombination pattern of Y chromosome and morphological difference of this chromosome between Bos taurus and Bos indicus make it an ideal model for studying structural variation, especially in crossbred （Bos taurus x Bos indicus） bulls. Copy Number Variation （CNV） is a type of genomic structural variation that gives information complementary to SNP data. The purpose of this study was to find out copy number differences of four Y chromosomal spermatogenesis-related candidate genes in genomic DNA of crossbred and purebred Indicine bulls. Result： Four Y chromosomal candidate genes of spermatogenesis namely, sex determining gene on Ychromosome （SRY）, DEAD box po/ypeptide 3-Y chromosome （DDX3 Y）, Ubiquidn specific peptidase 9, Y-linked （ usPgY）, testis-specific protein on Y chromosome （tspy） were evaluated. Absolute copy numbers of Y chromosomal genes were determined by standard curve-based quantitative real time PCR. Copy numbers of SRYand tspygenes per unit amount of genomic DNA are higher in crossbred than Indicine bulls. However, no difference was observed in DDX3Yand usPgYgene copy numbers between two groups. Conclusion： The present study demonstrates that the structural organization of Y chromosomes differs between crossbred and Indicine bulls which are reproductively healthy as observed from analysis of semen attributes. The absolute copy numbers of SRY and tspy genes in unit mass of genomic DNA of crossbred bulls are significantly higher than Indicine bulls. No alteration in absolute copies ofDDX3Yand usPgYgene was found between the genome of crossbred and Indicine bulls. This study suggests that the DDX3Yand USPgYare likely to be single copy genes in the genome of crossbred and Indicine bulls and variation in Y chromosome length between crossbred and Indicine bulls may be due to the copy number variation of

关键词： Absolute copy SRY DDX3Y usPgY tspy Crossbred

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绵羊Y染色体基因结构和表达特征及多拷贝基因CNV研究

绵羊Y染色体基因结构和表达特征及多拷贝基因CNV研究

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作者：徐海月兰州大学

学位级别：硕士

哺乳动物Y染色体雄性特异区基因主要或者特异在睾丸中表达,与睾丸发育紧密相关。Y染色基因结构复杂,具有物种特异性。绵羊Y染色体序列草图虽然已经发布,但是其包含基因的结构和表达特征、多拷贝基因的拷贝数变异(Copy number variation,... 详细信息

哺乳动物Y染色体雄性特异区基因主要或者特异在睾丸中表达,与睾丸发育紧密相关。Y染色基因结构复杂,具有物种特异性。绵羊Y染色体序列草图虽然已经发布,但是其包含基因的结构和表达特征、多拷贝基因的拷贝数变异(Copy number variation,CNV)还有待深入研究。本论文基于湖羊不同月龄,包括0月龄(初生)、3月龄(初情期)、6月龄(性成熟)、12月龄(体成熟)睾丸组织的二代转录组和不同月龄睾丸组织混合样的三代转录组数据,并以NCBI公布的湖羊Y染色体基因组(GCA＿011170295.1)和绵羊基因组(Oar＿rambouillet＿v1.0)为参考,精细注释绵羊Y染色体基因结构,分析Y染色体基因的表达特征,并利用q PCR技术验证相关基因的表达结果;同时利用q PCR技术确定两个多拷贝基因HSFY和tspy在不同品种中的CNV特征,分析CNV与湖羊睾丸大小性状的相关性,以期为Y染色体基因在绵羊育种中的利用提供科学依据。本论文主要研究结果如下:(1)通过多组学数据优化原有的基因注释模型,在绵羊Y染色体上注释出10个单拷贝基因,获得各基因详细的基因结构。基于不同月龄睾丸的RNA-Seq数据,获得10个基因在湖羊睾丸不同发育阶段的表达特征,其中DDX3Y、ZRSR2Y、OFD1Y基因的表达量随月龄的升高而逐渐增加。在睾丸不同发育阶段中,单拷贝基因AMELY、RBMY、ZFY、UTY、SRY、EIF2S3Y和USP9Y的表达并没有表现出时序特征。(2)注释出4个多拷贝基因,包括tspy、ZNF280BY、HSFY、PRAMEY基因,其中本研究首次注释出绵羊tspy基因。RNA-Seq数据显示4个基因的表达量均随月龄的升高而增加,其中在3月龄至6月龄表达显著变化。q PCR检测发现HSFY和tspy主要在睾丸中表达,并且表达量极显著高于心脏、肝脏、脾脏、肺脏、肾脏、肌肉组织。(3)利用q PCR对8个品种492只绵羊的tspy和HSFY基因进行CNV研究,发现tspy和HSFY基因在8个品种绵羊中均存在拷贝数变异。tspy的中值拷贝数为57(范围2-207),南非肉用美利奴羊(77,范围9-152)和杜泊羊(77,范围3-120)中值拷贝数最高,而滩羊中值拷贝数最低(20,范围10-92)。HSFY的中值拷贝数为5(范围1-24),南非肉用美利奴羊(7,范围3-12)中值拷贝数最高,滩羊最低(1,范围1-6)。tspy基因的CNV与部分睾丸性状呈显著相关。

关键词： Y染色体 HSFY tspy 基因表达拷贝数变异

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