Azure mutants: a type of host-dependent mutant of the bacteriophage f2

A new type of host-dependent mutant, azure mutant, of bacteriophage f2 has been isolated. Growth of these mutants was restricted specifically by amber suppressor genes in the host bacteria. Restriction of the formation of infective centers by different bacterial suppressor genes was 98 percent, 90 percent, and 70 percent with Su-3, Su-1, and Su-2 genes, respectively. Restriction, like suppression, was the dominant phenotype. The block in growth of the mutants occurred in an early stage of the infection cycle. Once infection was established, however, an infected cell produced approximately the same number of progeny phage as a cell without the suppressor genes did. It is proposed that the azure codon is the same as the amber codon (uracil, adenine, guanine) and that restriction results from improper termination of protein chains of the phage RNA polymerase. Similar mutants may exist in other systems.     

The approaching era of the tumor suppressor genes

Genes that can inhibit the expression of the tumorigenic phenotype have been detected by the fusion of normal and malignant cells, the phenotypic reversion of in vitro transformants, the induction of terminal differentiation of malignant cell lineages, the loss of "recessive cancer genes," the discovery of regulatory sequences in the immediate vicinity of certain oncogenes, and the inhibition of tumor growth by normal cell products. Such tumor suppressor genes will probably turn out to be as, if not more, diversified as the oncogenes. Consideration of both kinds of genes may reveal common or interrelated functional properties.     

Cooperative regulation of cell polarity and growth by Drosophila tumor suppressors

Loss of cell polarity and tissue architecture are characteristics of malignant cancers derived from epithelial tissues. We provide evidence from Drosophila that a group of membrane-associated proteins act in concert to regulate both epithelial structure and cell proliferation. Scribble (Scrib) is a cell junction-localized protein required for polarization of embryonic and, as demonstrated here, imaginal disc and follicular epithelia. We show that the tumor suppressors lethal giant larvae (lgl) and discs-large (dlg) have identical effects on all three epithelia, and that scrib also acts as a tumor suppressor. Scrib and Dlg colocalize and overlap with Lgl in epithelia; activity of all three genes is required for cortical localization of Lgl and junctional localization of Scrib and Dlg. scrib, dlg, and lgl show strong genetic interactions. Our data indicate that the three tumor suppressors act together in a common pathway to regulate cell polarity and growth control.     

Regulation of cell migration by the C2 domain of the tumor suppressor PTEN

PTEN is a tumor suppressor protein that dephosphorylates phosphatidylinositol 3,4,5 trisphosphate and antagonizes the phosphatidylinositol-3 kinase signaling pathway. We show here that PTEN can also inhibit cell migration through its C2 domain, independent of its lipid phosphatase activity. This activity depends on the protein phosphatase activity of PTEN and on dephosphorylation at a single residue, threonine(383). The ability of PTEN to control cell migration through its C2 domain is likely to be an important feature of its tumor suppressor activity.     

The control of the metabolic switch in cancers by oncogenes and tumor suppressor genes

Cells from some tumors use an altered metabolic pattern compared with that of normal differentiated adult cells in the body. Tumor cells take up much more glucose and mainly process it through aerobic glycolysis, producing large quantities of secreted lactate with a lower use of oxidative phosphorylation that would generate more adenosine triphosphate (ATP), water, and carbon dioxide. This is the Warburg effect, which provides substrates for cell growth and division and free energy (ATP) from enhanced glucose use. This metabolic switch places the emphasis on producing intermediates for cell growth and division, and it is regulated by both oncogenes and tumor suppressor genes in a number of key cancer-producing pathways. Blocking these metabolic pathways or restoring these altered pathways could lead to a new approach in cancer treatments.     

Mutational analysis of the tyrosine phosphatome in colorectal cancers

Tyrosine phosphorylation, regulated by protein tyrosine phosphatases (PTPs) and kinases (PTKs), is important in signaling pathways underlying tumorigenesis. A mutational analysis of the tyrosine phosphatase gene superfamily in human cancers identified 83 somatic mutations in six PTPs (PTPRF, PTPRG, PTPRT, PTPN3, PTPN13, PTPN14), affecting 26% of colorectal cancers and a smaller fraction of lung, breast, and gastric cancers. Fifteen mutations were nonsense, frameshift, or splice-site alterations predicted to result in truncated proteins lacking phosphatase activity. Five missense mutations in the most commonly altered PTP (PTPRT) were biochemically examined and found to reduce phosphatase activity. Expression of wild-type but not a mutant PTPRT in human cancer cells inhibited cell growth. These observations suggest that the mutated tyrosine phosphatases are tumor suppressor genes, regulating cellular pathways that may be amenable to therapeutic intervention.     

抑制体细胞衰老促进诱导性多潜能干细胞（iPSC）生成的研究进展

诱导性多潜能干细胞（induced pluripotent stem cell, iPSC）是指对体细胞实施特定的诱导方法,将体细胞重编程获得的多潜能干细胞。最常用的诱导方法是利用基因导入技术,将与胚胎干细胞（embryonic stem cell, ESC）多潜能性相关基因的转录因子导入体细胞,激活内源多能性基因的表达,实现体细胞重编程。除转基因外,使用某些小分子物质或蛋白质等也可以实现体细胞重编程。iPSC与ESC在形态学、表观遗传学和分化能力上高度相似。由于iPSC来源于普通成体细胞,避免了胚胎干细胞面临的伦理道德和免疫排斥问题,使得这一技术在再生医学和畜牧生产上都具有广阔的应用前景。然而,iPSC的低生成率和高风险性逐渐成为制约该技术发展的主要障碍。生成效率和速率低,大大增加了获得iPSC的难度,工作量大且成本高;高风险性使iPSC无法安全应用于再生医学和生产转基因动物。这成为iPSC科研工作者亟待解决的两大问题。文章介绍了通过抑制体细胞衰老来促进iPSC生成的相关研究进展。该方法对体细胞重编程有显著效果,但同时也存在较大的安全性争议。控制细胞衰老凋亡的Ink4a/Arf位点,p53、pRB、p21等基因和蛋白因子可以及时清除体内损伤细胞,促进细胞衰老凋亡,抑制细胞癌变,组成了维护机体健康的重要调控通路。近年研究表明,抑制促细胞衰老凋亡基因的表达可显著提高iPSC生成的效率和速率,说明肿瘤发生和iPSC生成存在某些相同的调控通路。抑制体细胞衰老的方法主要有3类：改进培养液成分、使用新的转录因子和调节细胞培养环境。通过抑制体细胞衰老,最高可将小鼠iPSC生成效率提高到100%。这为高效获得iPSC,探索iPSC及肿瘤的生成机制提供了新思路。同时,此方法存在较大安全性问题。iPSC最初是由Oct4,Sox2,Klf4,c-Myc等4种转录因子诱导而来,这些因子本身都存在致癌风险。抑癌基因的失活使获得的iPSC致癌风险增大,成为制约该方法广泛应用的最主要障碍。文章概括了2008年以来通过抑制体细胞衰老促进iPSC生成的研究进展、存在问题和应用前景。     

Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells

Classical genetic screens can be limited by the selectivity of mutational targeting, the complexities of anatomically based phenotypic analysis, or difficulties in subsequent gene identification. Focusing on signaling response to the secreted morphogen Hedgehog (Hh), we used RNA interference (RNAi) and a quantitative cultured cell assay to systematically screen functional roles of all kinases and phosphatases, and subsequently 43% of predicted Drosophila genes. Two gene products reported to function in Wingless (Wg) signaling were identified as Hh pathway components: a cell surface protein (Dally-like protein) required for Hh signal reception, and casein kinase 1alpha, a candidate tumor suppressor that regulates basal activities of both Hh and Wg pathways. This type of cultured cell-based functional genomics approach may be useful in the systematic analysis of other biological processes.     

Tet-on系统诱导猿猴病毒40T在CHO细胞中的表达

四环素诱导表达系统（Tet-off／Tet-on系统）是比较成熟的真核生物基因诱导表达系统之一,具有高效、无毒、严密开／关功能的特点。猿猴病毒40T（SV40T）是一种病毒癌蛋白,其与肿瘤抑制蛋白p53和Rb结合,并使之失活,从而消除它们抑制细胞生长的功能,使细胞分裂加速,形成肿瘤。利用Tet-on系统首先稳定筛选获得了表达Tet-on系统调节元件rtTA的阳性细胞CHO-pTet-on,再通过稳定筛选又成功得到导入其反应元件的双阳性细胞CHO-pTet-on-pTRE2-SV40T-Hyg,经强力霉素诱导表达了目的基因SV40T,建立了Tet-on基因诱导表达系统的细胞诱导表达研究平台。     

Germ line p53 mutations in a familial syndrome of breast cancer, sarcomas, and other neoplasms

Familial cancer syndromes have helped to define the role of tumor suppressor genes in the development of cancer. The dominantly inherited Li-Fraumeni syndrome (LFS) is of particular interest because of the diversity of childhood and adult tumors that occur in affected individuals. The rarity and high mortality of LFS precluded formal linkage analysis. The alternative approach was to select the most plausible candidate gene. The tumor suppressor gene, p53, was studied because of previous indications that this gene is inactivated in the sporadic (nonfamilial) forms of most cancers that are associated with LFS. Germ line p53 mutations have been detected in all five LFS families analyzed. These mutations do not produce amounts of mutant p53 protein expected to exert a trans-dominant loss of function effect on wild-type p53 protein. The frequency of germ line p53 mutations can now be examined in additional families with LFS, and in other cancer patients and families with clinical features that might be attributed to the mutation.     

FBXW7 targets mTOR for degradation and cooperates with PTEN in tumor suppression

The enzyme mTOR (mammalian target of rapamycin) is a major target for therapeutic intervention to treat many human diseases, including cancer, but very little is known about the processes that control levels of mTOR protein. Here, we show that mTOR is targeted for ubiquitination and consequent degradation by binding to the tumor suppressor protein FBXW7. Human breast cancer cell lines and primary tumors showed a reciprocal relation between loss of FBXW7 and deletion or mutation of PTEN (phosphatase and tensin homolog), which also activates mTOR. Tumor cell lines harboring deletions or mutations in FBXW7 are particularly sensitive to rapamycin treatment, which suggests that loss of FBXW7 may be a biomarker for human cancers susceptible to treatment with inhibitors of the mTOR pathway.     

Externally suppressible proline quadruplet ccc U

Three (+1) frameshift mutations located at different genetic sites respond with high specificity to the same external suppressor. In each case, the suppressor restores small amounts of protein that is normal in electrophoretic mobility and heat stability. One of these proteins has been shown to have the wildtype amino acid sequence. The messenger RNA quadruplet CCCUappears to be common to all three frameshift sites and to be translated by the suppressor as proline. A likely suppressor agent is a proline transfer RNA with a quadruplet anticodon or its functional equivalent.     

Orientation of asymmetric stem cell division by the APC tumor suppressor and centrosome

Stem cell self-renewal can be specified by local signals from the surrounding microenvironment, or niche. However, the relation between the niche and the mechanisms that ensure the correct balance between stem cell self-renewal and differentiation is poorly understood. Here, we show that dividing Drosophila male germline stem cells use intracellular mechanisms involving centrosome function and cortically localized Adenomatous Polyposis Coli tumor suppressor protein to orient mitotic spindles perpendicular to the niche, ensuring a reliably asymmetric outcome in which one daughter cell remains in the niche and self-renews stem cell identity, whereas the other, displaced away, initiates differentiation.     

基因表达系列分析技术在植物基因表达分析中的应用

 基因表达系列分析技术 (SAGE)是一种以测序为基础 ,采用数字化分析手段 ,在转录物水平上研究细胞或组织基因表达模式的有效工具。该技术不仅能够全面地分析特定组织或细胞表达的基因 ,获得这些基因表达丰度的数量信息 ,还可比较不同组织、不同时空条件下基因表达的差异 ,从而发现新基因。SAGE技术在植物方面的应用相对较少 ,但进展很快。本文着重介绍了SAGE在植物基因表达分析中的应用 ,分析SAGE所存在的问题、改进方法及发展前景     

p53 dynamics control cell fate

Cells transmit information through molecular signals that often show complex dynamical patterns. The dynamic behavior of the tumor suppressor p53 varies depending on the stimulus; in response to double-strand DNA breaks, it shows a series of repeated pulses. Using a computational model, we identified a sequence of precisely timed drug additions that alter p53 pulses to instead produce a sustained p53 response. This leads to the expression of a different set of downstream genes and also alters cell fate: Cells that experience p53 pulses recover from DNA damage, whereas cells exposed to sustained p53 signaling frequently undergo senescence. Our results show that protein dynamics can be an important part of a signal, directly influencing cellular fate decisions.     

Senescence of nickel-transformed cells by an X chromosome: possible epigenetic control

Transfer of a normal Chinese hamster X chromosome (carried in a mouse A9 donor cell line) to a nickel-transformed Chinese hamster cell line with an Xq chromosome deletion resulted in senescense of these previously immortal cells. At early passages of the A9/CX donor cells, the hamster X chromosome was highly active, inducing senescence in 100% of the colonies obtained after its transfer into the nickel-transformed cells. However, senescence was reduced to 50% when Chinese hamster X chromosomes were transferred from later passage A9 cells. Full senescing activity of the intact hamster X chromosome was restored by treatment of the donor mouse cells with 5-azacytidine, which induced demethylation of DNA. These results suggest that a senescence gene or genes, which may be located on the Chinese hamster X chromosome, can be regulated by DNA methylation, and that escape from senescence and possibly loss of tumor suppressor gene activity can occur by epigenetic mechanisms.