Mantovani R, Pessara U, Tronche F, Li X Y, Knapp A M, Pasquali J L, Benoist C, Mathis D
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, France.
EMBO J. 1992 Sep;11(9):3315-22. doi: 10.1002/j.1460-2075.1992.tb05410.x.
NF-Y is a sequence-specific DNA-binding protein which, as a heterodimer, recognizes CCAAT motifs in a variety of transcriptional promoters. We have generated a panel of monoclonal and affinity-purified polyclonal antibodies directed against various epitopes of NF-Y. These reagents are highly specific for either of the A or B subunits; we have mapped the epitopes recognized by the monoclonal antibodies to the glutamine-rich activation domain of NF-YA. The antibodies inhibit in vitro transcription from the promoters of the albumin gene and of Ea, a class II gene of the major histocompatibility complex. These data definitively demonstrate the role of NF-Y in regulating the transcription of two tissue-specific genes whose expression patterns do not overlap. Interestingly, the antibodies cannot inhibit a formed pre-initiation complex, but do block reinitiation of subsequent rounds of transcription from the same templates.
核因子Y（NF-Y）是一种序列特异性DNA结合蛋白，它作为异二聚体，能识别多种转录启动子中的CCAAT基序。我们已经制备了一组针对NF-Y各种表位的单克隆抗体和亲和纯化的多克隆抗体。这些试剂对A或B亚基中的任何一个都具有高度特异性；我们已将单克隆抗体识别的表位定位到NF-YA富含谷氨酰胺的激活结构域。这些抗体抑制白蛋白基因启动子和主要组织相容性复合体II类基因Ea启动子的体外转录。这些数据明确证明了NF-Y在调控两个组织特异性基因转录中的作用，这两个基因的表达模式并不重叠。有趣的是，这些抗体不能抑制已形成的预起始复合物，但确实会阻止同一模板后续转录轮次的重新起始。
