Wang D, Stravopodis D, Teglund S, Kitazawa J, Ihle J N
Department of Biochemistry, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
Mol Cell Biol. 1996 Nov;16(11):6141-8. doi: 10.1128/MCB.16.11.6141.
Stat5 was initially identified as a prolactin-induced member of the signal transducer and activator of transcription (Stat) family in sheep. However, Stat5 is also activated in the response to a variety of cytokines. In mice, and possibly in other species, there exist two Stat5 genes (Stat5a and Stat5b) that encode proteins of 92 and 94 kDa that are 95% identical. In the studies described here, we demonstrate that naturally occurring carboxyl-truncated, variant Stat5 proteins of 77 and 80 kDa exist and that these proteins are inducibly tyrosine phosphorylated in the response to several cytokines and form heterodimers with the full-length, wild-type proteins. Using expression constructs encoding truncated forms, we demonstrate that the truncated forms can be tyrosine phosphorylated and bind DNA. Surprisingly, the tyrosine phosphorylation of the carboxyl-truncated forms is considerably more stable than that of the wild-type proteins. Overexpression of a carboxyl-truncated Stat5a in cells resulted in the specific inhibition of the transcriptional activation by interleukin-3 of the genes for oncostatin M (Osm) and the cytokine-inducible, SH2 domain-containing gene (Cis), both of which have been shown to be normally regulated by Stat5. Although Stat5 dominantly suppressed the induction of these genes, no effects on cell proliferation were observed. Together, the results demonstrate the natural existence of potentially dominantly suppressive variants of Stat5 and implicate the carboxyl domain of Stats in transcriptional regulation and functions related to dephosphorylation.
Stat5最初是作为绵羊中催乳素诱导的信号转导子和转录激活子(Stat)家族成员被鉴定出来的。然而,Stat5在对多种细胞因子的应答中也会被激活。在小鼠以及可能在其他物种中,存在两个Stat5基因(Stat5a和Stat5b),它们编码92 kDa和94 kDa的蛋白质,这两种蛋白质有95%的同源性。在本文所述的研究中,我们证明存在天然的羧基端截短的、77 kDa和80 kDa的变异Stat5蛋白,并且这些蛋白在对几种细胞因子的应答中可被诱导酪氨酸磷酸化,并与全长野生型蛋白形成异二聚体。使用编码截短形式的表达构建体,我们证明截短形式可以被酪氨酸磷酸化并结合DNA。令人惊讶的是,羧基端截短形式的酪氨酸磷酸化比野生型蛋白的酪氨酸磷酸化要稳定得多。在细胞中过表达羧基端截短的Stat5a导致抑癌素M(Osm)基因和细胞因子诱导的含SH2结构域基因(Cis)的白细胞介素-3转录激活受到特异性抑制,这两个基因已被证明通常受Stat5调控。尽管Stat5主要抑制这些基因的诱导,但未观察到对细胞增殖的影响。总之,这些结果证明了Stat5潜在的显性抑制变异体的天然存在,并表明Stat蛋白的羧基结构域在转录调控和与去磷酸化相关的功能中起作用。