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一组人类推定的淋巴细胞G0/G1转换基因包括与啮齿动物细胞因子和锌指蛋白编码基因同源的基因。

A set of human putative lymphocyte G0/G1 switch genes includes genes homologous to rodent cytokine and zinc finger protein-encoding genes.

作者信息

Siderovski D P, Blum S, Forsdyke R E, Forsdyke D R

机构信息

Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.

出版信息

DNA Cell Biol. 1990 Oct;9(8):579-87. doi: 10.1089/dna.1990.9.579.

Abstract

Lymphocyte G0/G1 switch genes (G0S genes) are potential oncogenes and may regulate, be regulated by, or be coordinately regulated with, latent lymphotropic viruses. To identify these genes, a cDNA library was prepared from blood mononuclear cells that had been cultured for 2 hr with a T-cell mitogen (lectin) and cycloheximide. Eight differentially hybridizing recombinants were characterized by RNA and DNA blotting and sequencing. One cDNA (G0S7) corresponded to the oncogene c-fos. Another cDNA (G0S19) was homologous (70%) to a cDNA encoding a murine inhibitor of stem cell proliferation (the cytokine MIP1 alpha) and, less closely, to other members of the "small inducible" secreted protein-encoding gene family. Whereas cDNA hybridization to genomic DNA blots indicated a small subfamily of G0S19 genes, simple patterns of bands indicated that most cDNAs, including G0S30 cDNA, corresponded to single-copy genes. The 3' noncoding sequence of G0S30 cDNA was homologous (87-89%) to the 3' noncoding sequences of certain rodent genes (NGFI-A, Krox24, EGR1) that encode zinc finger proteins (putative transcriptional regulators). This degree of evolutionary conservation suggests an important function for the 3' noncoding region. The 3' noncoding regions of some cDNAs contained the TTATTTAT (mRNA destabilization) element. The corresponding RNAs each formed doublets in agarose gels. Previous studies of c-fos RNA from HeLa cells indicate that this is due to cycloheximide-dependent stabilization of poly(A) tails. Our results reveal the power of cycloheximide enrichment in isolating what would appear to be significant low-abundance mRNAs.

摘要

淋巴细胞G0/G1转换基因(G0S基因)是潜在的癌基因,可能与潜伏性嗜淋巴细胞病毒相互调节或协同调节。为了鉴定这些基因,从经T细胞丝裂原(凝集素)和环己酰亚胺培养2小时的血液单核细胞中制备了cDNA文库。通过RNA和DNA印迹及测序对8个差异杂交重组体进行了表征。一个cDNA(G0S7)对应于癌基因c-fos。另一个cDNA(G0S19)与编码小鼠干细胞增殖抑制剂(细胞因子MIP1α)的cDNA同源性为70%,与“小诱导型”分泌蛋白编码基因家族的其他成员同源性较低。cDNA与基因组DNA印迹杂交表明G0S19基因有一个小亚家族,而条带的简单模式表明大多数cDNA,包括G0S30 cDNA,对应于单拷贝基因。G0S30 cDNA的3'非编码序列与某些编码锌指蛋白(推定的转录调节因子)的啮齿动物基因(NGFI-A、Krox24、EGR1)的3'非编码序列同源性为87-89%。这种程度的进化保守性表明3'非编码区具有重要功能。一些cDNA的3'非编码区含有TTATTTAT(mRNA不稳定)元件。相应的RNA在琼脂糖凝胶中均形成双链体。先前对HeLa细胞中c-fos RNA的研究表明,这是由于环己酰亚胺依赖的多聚腺苷酸尾巴稳定化所致。我们的结果揭示了环己酰亚胺富集在分离看似重要的低丰度mRNA方面的作用。

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