Identification of JAK/STAT signalling components by genome-wide RNA interference
Identification of JAK/STAT signalling components by genome-wide RNA interference
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DOI：
10.1038/nature03869
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发表时间：
2005-08-11
期刊：
影响因子：
64.8
通讯作者：
Boutros, M
中科院分区：
文献类型：
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作者：
Müller, P;Kuttenkeuler, D;Boutros, M
介导细胞间信息转导的信号通路对细胞发育、细胞分化和体内平衡至关重要(1)。它们的失调也经常与人类恶性肿瘤有关。Janus酪氨酸激酶/信号换能器和转录激活器（JAK/STAT）通路代表了这样一个信号级联，其进化保守的作用包括细胞增殖和造血(2)。在这里，我们描述了JAK/STAT通路活性所需基因的系统全基因组调查。对培养的黑胃果蝇血细胞样细胞20,026种RNA干扰（RNAi）诱导的表型进行分析，鉴定出4种已知蛋白和86种以前未表征的蛋白编码相互作用基因。随后，基于细胞的上位实验用于根据这些蛋白与信号级联已知组分的相互作用对其进行分类。除了多种人类疾病基因同源物外，我们还发现酪氨酸磷酸酶Ptp61F和BRWD3的果蝇同源物，BRWD3是一种含溴结构域的蛋白，在白血病中被破坏(3)。此外，体内分析表明，被破坏的dBRWD3和过表达的Ptp61F作为白血病样血细胞肿瘤的抑制因子。该筛选代表了JAK/STAT信号所需的新位点的全面鉴定，并提供了与人类癌症相关的重要途径的分子见解。已鉴定的途径修饰物的人类同源物可能构成治疗干预的靶标。
Signalling pathways mediating the transduction of information between cells are essential for development, cellular differentiation and homeostasis(1). Their dysregulation is also frequently associated with human malignancies. The Janus tyrosine kinase/ signal transducer and activator of transcription (JAK/STAT) pathway represents one such signalling cascade whose evolutionarily conserved roles include cell proliferation and haematopoiesis(2). Here we describe a systematic genome-wide survey for genes required for JAK/STAT pathway activity. Analysis of 20,026 RNA interference (RNAi)-induced phenotypes in cultured Drosophila melanogaster haemocyte-like cells identified interacting genes encoding 4 known and 86 previously uncharacterized proteins. Subsequently, cell-based epistasis experiments were used to classify these proteins on the basis of their interaction with known components of the signalling cascade. In addition to multiple human disease gene homologues, we have found the tyrosine phosphatase Ptp61F and the Drosophila homologue of BRWD3, a bromo-domain-containing protein disrupted in leukaemia(3). Moreover, in vivo analysis demonstrates that disrupted dBRWD3 and overexpressed Ptp61F function as suppressors of leukaemia-like blood cell tumours. This screen represents a comprehensive identification of novel loci required for JAK/STAT signalling and provides molecular insights into an important pathway relevant for human cancer. Human homologues of identified pathway modifiers may constitute targets for therapeutic interventions.