Okada H, Watanabe T, Niki M, Takano H, Chiba N, Yanai N, Tani K, Hibino H, Asano S, Mucenski M L, Ito Y, Noda T, Satake M
Department of Cell Biology, Cancer Institute, Tokyo, Japan.
Oncogene. 1998 Nov 5;17(18):2287-93. doi: 10.1038/sj.onc.1202151.
The AML1 and PEBP2beta/CBFbeta genes encode the DNA-binding and non-binding subunits, respectively, of the heterodimeric transcription factor, PEBP2/CBF. Targeting each gene results in an almost identical phenotype, namely the complete lack of definitive hematopoiesis in the fetal liver on embryonic day 11.5 (E11.5). We examined and compared the expression levels of various hematopoiesis-related genes in wild type embryos and in embryos mutated for AML1 or PEBP2beta/CBFbeta. The RNAs were prepared from the yolk sacs of E9.5 embryos, from the aorta-gonad- mesonephros regions of E11.5 embryos and from the livers of E11.5 embryos and RT-PCR was performed to detect various gene transcripts. Transcripts were detected for most of the hematopoiesis-related genes that encode transcription factors, cytokines and cytokine receptors, even in tissues from homozygously targeted embryos. On the other hand, PU.1 transcripts were never detected in any tissue of AML1(-/-) or PEBP2beta/CBFbeta(-/-) embryos. In addition, transcripts for the Vav, flk-2/flt-3, M-CSF receptor, G-CSF receptor and c-Myb genes were not detected in certain tissues of the (-/-) embryos. The results suggest that the expression of a particular set of hematopoiesis-related genes is closely correlated with the PEBP2/CBF function.
AML1基因和PEBP2β/CBFβ基因分别编码异二聚体转录因子PEBP2/CBF的DNA结合亚基和非结合亚基。对每个基因进行靶向操作会导致几乎相同的表型,即在胚胎第11.5天(E11.5)时胎儿肝脏中完全缺乏定型造血。我们检测并比较了野生型胚胎以及AML1或PEBP2β/CBFβ基因发生突变的胚胎中各种造血相关基因的表达水平。RNA取自E9.5胚胎的卵黄囊、E11.5胚胎的主动脉-性腺-中肾区域以及E11.5胚胎的肝脏,并进行逆转录聚合酶链反应(RT-PCR)以检测各种基因转录本。即使在纯合靶向胚胎的组织中,也检测到了大多数编码转录因子、细胞因子和细胞因子受体的造血相关基因的转录本。另一方面,在AML1(-/-)或PEBP2β/CBFβ(-/-)胚胎的任何组织中均未检测到PU.1转录本。此外,在(-/-)胚胎的某些组织中未检测到Vav、flk-2/flt-3、M-CSF受体、G-CSF受体和c-Myb基因的转录本。结果表明,一组特定的造血相关基因的表达与PEBP2/CBF的功能密切相关。