◇◇新语丝(www.xys.org)(xys.dxiong.com)(xys.3322.org)(xys.freedns.us)◇◇ (中文部分为学生翻译,有不准确之处,请以后面的英文原文为准) 我们在清华大学的工作进展 傅新元   目前网上有一些关于清华的讨论。我感觉有必要澄清一下事实,并对我们已 经取得的成绩做一个简要的描述。   从2000年10月开始,我离开耶鲁大学在清华大学全职工作一年。其时,清华 学校投入了1000万元人民币建立基因组研究所。其中大约70%的资金被用于实验 室的改建和实验设备的购置(包括一套贵重的购自意大利的SPF级(三级)实验动 物设备在内)。由于此前清华大学缺乏分子生物学和实验动物领域相关的人员和 专门技术,所以我们在清华大学的全部工作几乎都是从零开始的。   剩余的约300万元人民币大致相当于在美国任意一所相当好水平的大学一个 新晋教员维持日常研究所需的启动经费。所不同的是,我们开展的是全新的工作, 而不是像美国的博士后初任助理教授那样,可以延续以前实验室的工作。另一个 因素是我们也需要从头开始训练学生,而不像在耶鲁的实验室里,新来的博士后 一开始就能独立的进行实验。但是,无论是在清华还是在美国,都需要至少两到 三年的积累才能看到成果。以上所述就是我们当时面临的情况。   一般来说,在分子生物学领域发表一篇好的文章需要几年的时间,从头开始 建立和完善一个新的系统应该需要更长的时间。以我个人为例,从1996年开始, 我们在耶鲁开展了一个研究STAT3功能的项目,直到7年后,才发表了一系列重要 的文章。(参见PNAS 100. 1879-1884. PNAS 100, 12929-12934; J. Experimental Med, 198, 1517-1525; Immunity, 19, 849?61)。我认为任何一 个分子生物学领域的项目负责人(PI)都会认可我的观点。   基于以上的评判标准和背景材料,我对于我们在清华的工作给予以下的评价。   在2003年,我们已经在JBC上发表了一篇研究hSef基因影响细胞分化功能的 文章,这篇文章最重要的作者就是常智杰老师。另外,一篇关于Wnt信号通路的 文章即将发表在享有盛誉的Molecular Cell杂志上,这项工作主要是在哈佛大学 贺熹教授的实验室完成的,我们实验室的常智杰老师和张新军同学作为共同作者 参加了这项工作。关于这个项目的最新进展我正在集中精力写一篇文章。   另外一个主要的进展是在清华独立完成的,那是一个关于TGF-beta-Smad 信 号通路新的调控机制的发现。这项工作是由常智杰老师和我们的学生在清华完成 的。(摘要附后)   这篇文章很快将会发表在分子生物学领域具有前沿地位的Mol. & Cell. Biol. (MCB,Vol.24,No.2 Jan 2004 issue)上。这是清华大学第一篇MCB文章。我想任 何分子生物学领域的人都应该知道MCB的影响力。一般来说,只有做出深入的功 能性和机理性研究的工作才能被MCB接收。   在这篇文章中,常智杰老师等人首次揭示: CHIP蛋白可以调控SMAD蛋白的 降解,而SMAD蛋白是TGF-beta信号通路的关键介导者。   我需要指出的是:对国内实验室而言,克隆一个重要的新基因不足为奇。然 而,目前困难的是对此基因开展深入的功能研究和揭示相应的分子机制。常老师 文章的重要之处就在于其机理性和功能性研究。通过一系列设计严格的实验,常 老师等人首次阐明了CHIP可以调控TGF-beta-Smad信号通路。任何有细胞信号传 导领域背景知识的人都会认识到这篇文章是TGF-beta 信号通路研究领域中一个 重要的发现,并且会产生重大的影响。   由于清华刚刚开展分子生物学的研究,可想而知,在竞争极其激烈的 TGF-beta 信号通路研究领域首次获得承认,我们必须付出更加艰苦的努力。从 文章写作,到按照审稿人的意见修改,到最后被MCB接收,常老师花了几乎一年 的时间。我必须声明,这仅仅是常老师取得成功的非常重要的第一步,我相信在 新的一年里,常老师和清华我们组的同学们能够发表更多更好的文章。如果有人 对此文章感兴趣,我将会提供此文的PDF格式预印本。   另外一个由刘力老师和我们清华的学生们开展的重要工作是一个新的癌基因 NOK的分子克隆与功能鉴定。从用生物信息学的方法克隆直到用动物肿瘤模型进 行功能性研究,都是在清华大学我们的实验室开展的。此研究最重要的发现是揭 示了NOK基因快速广泛的促肿瘤转移能力。我们已经建立了严格的动物模型,为 进一步研究这一新的受体-癌基因诱导肿瘤转移的内在机理打下了基础。癌症领 域的顶极杂志CANCER RESEARCH的审稿人对该文给予了乐观的评价,我们期望它 很快就能发表。   同时,我们很快就要投送更多的文章,其中包括:一个关于WT1肿瘤基因及 其相关蛋白新的重要发现;关于TGF-beta-Smad信号通路的第二篇文章;一篇关 于调控Wnt信号通路蛋白的突破性文章;一篇关于糖皮质激素受体调控机制的文章; 我们预期,在2004年,我们将投送至少5-7篇JBC或MCB水平的文章。   一旦实验进展中有苗头,我们将会瞄准更高的目标。我相信,在不久的将来, 我们实验室的科研成果将达到与我们有类似规模和资金支持的美国大学先进实验 室的水平。这也是我们建立实验室最初的目标。   总之,根据以上信息,每个科学家都能用他/她的良知去评判我们的工作进 展。但是对我而言,我有各种理由对我们在清华的工作感到自豪和乐观。无论用 各种标准评判,从零做起,在短短的三年里能取得现在的成果都是一个很好的开 始。 Our Progress in Tsinghua University Written by Xin-Yuan Fu, Ph. D. Dec. 25, 2003 in Tsinghua University There are some discussions in the website about our work in Tsinghua. I feel that it is necessary for me to clarify some facts and give a brief description what we have achieved in Tsinghua University. Tsinghua University has provide a fund about 10,000,000 Yuan (RMB) to the Tsinghua Institute of Genome Research, which was founded in Oct. 2000, the year I took a sabbatical leave from Yale to work in Tsinghua for a full year. About 70% of fund was used for remodel our laboratory and setting-up basic facilities, including an expensive pathogen-free animal facility imported from Italy. Since Tsinghua had little expertise and resources for molecular biology and animal work before we came, we basically started almost everything from "ground zero". The fund of about 3,000,000 RMB supporting for day-to-day research is about the same size as a start-up fund for a junior faculty member starting a laboratory in a decent University in USA. One difference is that all of our projects are completely new and started from very beginning, which is not like a continuation of work when a postdoctoral fellow becomes an assistant professor with carry-overs from previous lab in USA. Another factor is that we were training our students from very beginning too. I had well-trained and capable postdoctoral fellows in my lab at Yale, who can perform skilled experiments independently from the day one. However, any new students of molecular biology, no matter you are in USA or Tsinghua, may need at least two or three years experience to be productive. This is our situation in Tsinghua when we first started. In general, it took several years of work to get a good paper published in the molecular biology field. To develop and establish your own new system of research from very beginning could take longer time. For my personal experience, we started a project on STAT3 function in 1996 at Yale, and we had worked almost 7 years to get several significant papers published in 2003 (see PNAS 100. 1879-1884. PNAS 100, 12929-12934; J. Experimental Med, 198, 1517- 1525; Immunity, 19, 849?861). I expect that any one, who has some experiences as a PI in the research field of molecular biology, will likely agree with me. With the above standard of comparison and the background information, I would give my account of our work in Tsinghua. In the year of 2003, there is already a JBC paper (J. Biol. Chem., Vol. 278, Issue 50, 50273-50282, 2003) published from our group on a gene hSef and its function in inducing cell differentiation. The senior author of the paper is Prof. Chang Zhijie. Additionally, a paper on Wnt signaling, co-authored by Prof. Chang Zhijie and Zhang Xin-Jun of our group with the major work done in the lab of Prof He Xi of Harvard will be published soon in Molecular Cell, an esteemed Journal in molecular biology. In the following part, however, I will concentrate my description on some new progress of ours. One major progress we made completely and independently in Tsinghua, was a discovery about a novel mechanism in regulation of TGF-b-Smad signaling. This work was achieved by Professor Chang Zhijie and our students in Tsinghua (abstract listed below). This paper will be soon published in Mol. & Cell. Biol. (MCB, Vol.24, No.2 Jan 2004 issue), a leading Journal in molecular biology field. This is Tsinghua's first MCB paper. I think that every one who has studied molecular biology, should know the impact of MCB. Generally, you have to have functional and mechanistic studies to get papers in MCB. In this paper, Prof. Chang et al has shown, for the first time, a protein termed CHIP, regulates degradation of SMAD proteins, which are the key mediator for TGF-b signaling. Prof. Chang et al., independently identified CHIP protein, and found it could interact with SMAD. I should point out that it is not unusual for a laboratory in China to clone novel and important genes. However, the tough task now is to carry out in-depth functional studies and reveal possible molecular mechanisms by which the gene is involved. The highlights of Prof. Chang's paper are its mechanistic and functional studies. Through a series of well-designed experiments, Prof. Chang et al. have demonstrated for the first time that CHIP regulates TGF-b-Smad signaling. Anyone with current knowledge of signal transduction should recognize that this paper is an important discovery in the field of TGF-b signaling and will have significant impact. Since Tsinghua is a new player in molecular biology field, it is anticipated that we have to struggle much harder to get recognition for the first time in this extremely competitive field of TGF-b signaling. It took almost one year for Prof. Chang, from writing the paper, revising according to reviewer's tough comments, and to the final accepting of the paper in MCB. I would claim that, this is an important first success for Professor Chang Zhijie, and I expect that he and students in our group in Tsinghua will have many more to come in the next year. If anyone interested in this paper, I can send him/her a preprint in PDF. The second major story developed by Prof. Liu Li and our students in Tsinghua is about the identification and characterization of a novel oncogene NOK (see abstract below). All the work, from identification and cloning this gene using bioinformatic/genomic method, to functional studies using animal tumor models, were completely carried out in our laboratory in Tsinghua. The most dramatic finding of this study is to reveal the role of NOK in inducing rapid and wide- spread metastasis of the tumor. We have established a critical animal model for further studies of hidden-mechanisms of metastasis induced by a novel receptor- oncogene. This paper has been reviewed positively in Cancer Research, a top Journal in the cancer field and we expect its publication soon. At the same time, we are about to submit several more papers very soon, including one on an important new findings about WT1 tumor genes and its partner proteins; the second paper on TGF-b-SMAD signaling, a ground-breaking paper on a novel signaling protein for regulation of Wnt signaling, and a paper on an interesting story of a regulatory protein in glucocorticoid receptor (GR) function. We expect that we will have at least five to seven major papers to be submitted for publication in 2004 in well-established journals like MCB and JBC. We would aim higher whenever a possibility is given with our progress. I am confident that, in the near future the productivity of our lab will be definitely comparable with many leading laboratories in the universities in USA, which have the similar size and support. This was essentially my objective when we started work in Tsinghua. In summary, every scientist can judge our progress with his/her honesty and the above information. But for me, I have all the reasons to be proud and optimistic about our work in Tsinghua. In any standard, it is a sound start for a laboratory that started essentially from "ground zero" and has only three years of age. =================== CHIP MEDIATES DEGRADATION OF SMAD PROTEINS AND POTENTIALLY REGULATES SMAD- INDUCED TRANSCRIPTION Linyu Li1*, Hong Xin1*, Xialian Xu1, Mei Huang1, Xinjun Zhang1, Yue Chen1, Shuping Zhang1, Xin-Yuan Fu1,3# , and Zhijie Chang1# 1 Tsinghua Institute of Genome Research, Department of Biological Sciences and Biotechnology and School of Medicine, Tsinghua University, Beijing(100084), China 2 Department of Pathology, Yale University School of Medicine, New Haven, CT 06520, USA ABSTRACT TGF-/BMP family ligands interact with specific membrane receptor complexes that have serine/threonine kinase activities. The receptor phosphorylation and activation induced by the ligands leads to phosphorylation of the Smad proteins that translocate to the nucleus, controlling gene expression. Thus, regulation of Smad proteins is a key step in TGF-/BMP-induced signal transduction. Here we report a novel mechanism of the regulation of SMAD-mediated signaling, by which the Smad1 protein level is controlled through expression of the CHIP protein. CHIP is a U-box-dependent E3 ubiquitin ligase, previously identified as a co-chaperon protein. However, we have isolated CHIP as a Smad-interacting protein in a yeast two-hybrid screen using Smad1 as bait. Furthermore we have shown CHIP/Smad interaction using the 35S-labeled CHIP protein that can interact with GST-Smad1 and GST-Smad4 in an in vitro protein-binding assay. The CHIP/Smad interaction has been confirmed in vivo in mammalian cells through co- immunoprecipitation. Interestingly, we demonstrate that the co-expression of Smad1 and Smad4 with the CHIP protein results in the degradation of the Smad proteins through a ubiquitin-mediated process. Consistent with the observation that CHIP induces Smad1 degradation, we further show that the expression of CHIP can inhibit the transcriptional activities of the Smad1/4 complex induced by BMP signals. Intriguingly, pBS/U6/CHIPi, which diminishes CHIP expression, significantly enhanced Smad1/4 or BMPRIB(QD) induced gene transcription. These results suggest that CHIP can interact with Smad1/4 proteins and block BMP signal transduction through the ubiquitin-mediated degradation of Smad proteins. A Novel Protein Tyrosine Kinase NOK that Shares Homology with PDGF/FGF Receptors, Induces Tumorigenesis and Metastasis in Nude Mice1 Li Liu2, Xin-Zi Yu, Tie-Shi Li, Lianxia Song, Pei-La Chen, Talin Suo, Ying-Hua Li, Shidong Wang, Yue Chen, Yongming Ren, Shuping Zhang, Zhijie Chang, Xin-Yuan Fu2 Tsinghua Institute of Genome Research, Department of Biological Sciences and Biotechnology [L.L., X.Z.Y., T.S.L., L.S., P.L.C., T.S., Y.H.L., S.D., Y.C.,Y.R., S.Z., Z.J, X.Y.F.], Peking Union Medical College [L.L.], Tsinghua University, Beijing 100084, China Department of Pathology, Yale University School of Medicine, New Haven, CT, USA [X.Y.F.] ABSTRACT Receptor protein tyrosine kinases (RPTKs) play important roles in regulation of a variety of cellular processes including cell migration, proliferation, and protection from apoptosis. Here we report the identification and characterization of a novel RPTK-like molecule that has a critical role in induction of tumorigenesis and metastasis and is termed NOK (novel oncogene with kinase-domain). NOK contains a putative single transmembrane domain and a conserved intracellular tyrosine kinase domain that shares homology with members of the PDGF/FGF receptor superfamily. NOK is exclusively located in the cytoplasm. NOK mRNAs are present in limited human organs and expressed with the highest abundance in prostate. A variety of tumor cells also express the NOK mRNAs. In this report, we demonstrate that NIH3T3 and BaF3 cells are strongly transformed by expression of NOK gene as examined by colony formation experiments. Furthermore, BaF3 cells with stable expression of NOK induced rapid tumorigenesis in nude mice. Interestingly, these NOK expressing tumor cells could promptly invade and spread into various distinct organs and form metastatic foci, eventually leading to rapid death of these animal. Thus, our results both in vitro and in vivo, suggest that NOK is a novel oncogene with capacity of promoting cell transformation, tumorigenesis and metastasis. (XYS20031226) ◇◇新语丝(www.xys.org)(xys.dxiong.com)(xys.3322.org)(xys.freedns.us)◇◇