Marchetti Marcella, Fanti Laura, Berloco Maria, Pimpinelli Sergio
Istituto Pasteur, Fondazione Cenci Bolognetti, Dipartimento di Genetica e Biologia Molecolare, Università La Sapienza, 00185 Rome, Italy.
Development. 2003 Aug;130(16):3683-9. doi: 10.1242/dev.00587.
We have analyzed the expression of homeotic Bithorax Complex proteins in the fat bodies of Drosophila larvae by staining with specific antibodies. We have found that these proteins are differentially expressed along the anteroposterior (AP) axis of the fat body, with patterns parallel to those previously characterized for the larval and adult epidermis. As fat body nuclei have polytene chromosomes, we were able to identify the BX-C locus and show that it assumes a strongly puffed conformation in cells actively expressing the genes of the BX-C. Immunostaining of these polytene chromosomes provided the resolution to cytologically map binding sites of the three proteins: Ubx, Abd-A and Abd-B. The results of this work provide a system with which to study the positioning of chromatin regulatory proteins in either a repressed and/or active BXC at the cytological level. In addition, the results of this work provide a map of homeotic target loci and thus constitute the basis for a systematic identification of genes that are direct in vivo targets of the BX-C genes.
我们通过用特异性抗体染色,分析了果蝇幼虫脂肪体中同源异形双胸复合体蛋白的表达情况。我们发现这些蛋白沿脂肪体的前后轴呈差异表达,其模式与先前在幼虫和成虫表皮中所描述的模式相似。由于脂肪体细胞核具有多线染色体,我们能够鉴定出BX-C基因座,并表明它在积极表达BX-C基因的细胞中呈现出强烈的胀泡构象。对这些多线染色体进行免疫染色,为在细胞学水平上绘制三种蛋白(Ubx、Abd-A和Abd-B)的结合位点提供了分辨率。这项工作的结果提供了一个系统,可用于在细胞学水平上研究染色质调节蛋白在被抑制和/或活跃的BXC中的定位。此外,这项工作的结果提供了一个同源异形靶基因座图谱,从而构成了系统鉴定作为BX-C基因直接体内靶标的基因的基础。