In vivo binding pattern of a trans-regulator of homoeotic genes in Drosophila melanogaster

The specification and maintenance of the metameric pattern in Drosophila melanogaster is regulated by complicated gene interactions. The differential expression of the homoeotic genes of the Antennapedia complex (ANT-C) and bithorax complex (BX-C), which determine segmental identities, is partly controlled by cross-regulatory interactions of loci within the two clusters and partly by trans-acting factors located outside the two complexes. One of the trans-regulatory genes, Polycomb (Pc), acts as a repressor of the ANT-C and BX-C. Mutations of Polycomb result in a complete depression of the homoeotic genes, leading to abdominal transformations of all body segments. Polycomb is part of a large class of trans-regulatory genes (Pc-group), estimated to comprise up to 40 loci. We have raised antibodies against the Polycomb protein, and, using an improved immunostaining technique, showed that the Polycomb protein binds to 60 discrete sites along the polytene chromosomes of salivary glands. These sites comprise the ANT-C and the BX-C as well as several locations of Pc-group genes. This is the first clear evidence for a direct interaction of Polycomb with homoeotic loci and other Pc-group genes.     

Deformed autoregulatory element from Drosophila functions in a conserved manner in transgenic mice.

The striking similarities in the structure, organization and anterior-posterior expression patterns between the murine Hox gene system and the Drosophila homeotic gene complexes, called HOM-C (ref. 3), may point to highly conserved mechanisms for specifying positional identities (reviewed in ref. 4). Strong support for this concept lies in the observation of conserved colinearity between the genomic order of the Hox/HOM genes and their unique successive expression domains along the anterior-posterior axes of both mouse and fly embryos. These unique and precise expression patterns appear to be facilitated by multiple cis-regulatory elements (reviewed in ref. 5). One of the few elements characterized in detail is the autoregulatory enhancer of the homeotic gene Deformed (Dfd), which supports expression in subregions of posterior head segments of Drosophila embryos. Here we present evidence that this enhancer is capable of conferring reporter gene expression to a discrete subregion of the hindbrain in transgenic mouse embryos. Remarkably, this anterior-posterior subregion lies within the common anterior expression domain of the Dfd cognate Hox genes in the postotic hindbrain. Our results indicate that the Dfd autoregulatory enhancer is part of a highly conserved mechanism for establishing region-specific gene expression along the anterior-posterior axis of the embryo.     

Homeotic transformation of the occipital bones of the skull by ectopic expression of a homeobox gene.

Murine Hox genes have been postulated to play a role in patterning of the embryonic body plan. Gene disruption studies have suggested that for a given Hox complex, patterning of cell identity along the antero-posterior axis is directed by the more 'posterior' (having a more posterior rostral boundary of expression) Hox proteins expressed in a given cell. This supports the 'posterior prevalence' model, which also predicts that ectopic expression of a given Hox gene would result in altered structure only in regions anterior to its normal domain of expression. To test this model further, we have expressed the Hox-4.2 gene more rostrally than its normal mesoderm anterior boundary of expression, which is at the level of the first cervical somites. This ectopic expression results in a homeotic transformation of the occipital bones towards a more posterior phenotype into structures that resemble cervical vertebrae, whereas it has no effect in regions that normally express Hox-4.2. These results are similar to the homeotic posteriorization phenomenon generated in Drosophila by ectopic expression of genes of the homeotic complex HOM-C (refs 7-10; reviewed in ref. 3).     

Activation of a C. elegans Antennapedia homologue in migrating cells controls their direction of migration.

Anterior-posterior patterning in insects, vertebrates and nematodes involves members of conserved Antennapedia-class homeobox gene clusters (HOM-C) that are thought to give specific body regions their identities. The effects of these genes on region-specific body structures have been described extensively, particularly in Drosophila, but little is known about how HOM-C genes affect the behaviours of cells that migrate into their domains of function. In Caenorhabditis elegans, the Antennapedia-like HOM-C gene mab-5 not only specifies postembryonic fates of cells in a posterior body region, but also influences the migration of mesodermal and neural cells that move through this region. Here we show that as one neuroblast migrates into this posterior region, it switches on mab-5 gene expression; mab-5 then acts as a developmental switch to control the migratory behaviour of the neuroblast descendants. HOM-C genes can therefore not only direct region-specific patterns of cell division and differentiation, but can also act within migrating cells to programme region-specific migratory behaviour.     

Domain of Ultrabithorax expression in Drosophila visceral mesoderm from autoregulation and exclusion

Domains of differential homeotic gene activity are formed at specific positions along the anteroposterior axis of the early Drosophila embryo. Homeotic genes are required continuously throughout development, so that homeotic gene activity has to be maintained independently of the positional information provided in the early embryo. In the ectoderm, the domains of homeotic gene activity partially overlap, but we have found that in the visceral mesoderm at least three of these genes are expressed in adjacent and mutually exclusive domains. It has been proposed that stable, sharply demarcated domains of this type could be established if a homeotic gene product stimulated its own expression locally and inhibited the expression of other homeotic genes, which Meinhardt has termed autocatalysis and mutual exclusion respectively. Furthermore, autocatalysis of this kind can in principle account for the maintenance of homeotic gene activity throughout development. We find that the unique domain of Ultrabithorax (Ubx) expression in the visceral mesoderm is dependent both on autocatalysis and on an exclusion mechanism: Ubx product is required for its own synthesis, whereas the product of the posteriorly adjacent gene abdominal-A represses Ubx expression.     

Isolation of a homoeo box-containing gene from the engrailed region of Drosophila and the spatial distribution of its transcripts

The engrailed locus of Drosophila melanogaster has the characteristics of both a homoeotic gene and a segmentation gene: like a homoeotic gene, it specifies the development of specific compartments of the Drosophila embryo (the posterior compartments of each segment), and, like mutations of segmentation genes, lethal alleles of engrailed affect also the pattern of segmentation of the embryo. Here we report that like many of the homoeotic genes of the bithorax and Antennapedia complexes, engrailed has a 'homoeo box' sequence: also, like the segmentation gene fushi tarazu, the engrailed gene displays a periodic pattern of expression in Drosophila embryos.     

Ultrabithorax is required for membranous wing identity in the beetle Tribolium castaneum

The two pairs of wings that are characteristic of ancestral pterygotes (winged insects) have often undergone evolutionary modification. In the fruitfly, Drosophila melanogaster, differences between the membranous forewings and the modified hindwings (halteres) depend on the Hox gene Ultrabithorax (Ubx). The Drosophila forewings develop without Hox input, while Ubx represses genes that are important for wing development, promoting haltere identity. However, the idea that Hox input is important to the morphologically specialized wing derivatives such as halteres, and not the more ancestral wings, requires examination in other insect orders. In beetles, such as Tribolium castaneum, it is the forewings that are modified (to form elytra), while the hindwings retain a morphologically more ancestral identity. Here we show that in this beetle Ubx 'de-specializes' the hindwings, which are transformed to elytra when the gene is knocked down. We also show evidence that elytra result from a Hox-free state, despite their diverged morphology. Ubx function in the hindwing seems necessary for a change in the expression of spalt, iroquois and achaete-scute homologues from elytron-like to more typical wing-like patterns. This counteracting effect of Ubx in beetle hindwings represents a previously unknown mode of wing diversification in insects.     

A mouse gene related to Distal-less shows a restricted expression in the developing forebrain.

Many genes known to be involved in embryogenesis and morphogenesis of the fruitfly Drosophila melanogaster encode proteins with a highly conserved region of 60 amino acids called the homeodomain. Mammalian counterparts for most of these genes have been identified, including those homologous to the Drosophila homeotic genes or to genes such as evenskipped, engrailed or caudal. We have isolated a murine homeobox gene that encodes a homeodomain similar to that encoded by the Drosophila Distalless (Dll) gene. Dll has a crucial role in Drosophila limb morphogenesis, partially specifying pattern along the proximo-distal axis of the limb. The murine counterpart is expressed in a restricted region of the developing brain, within the diencephalon and the adjacent telencephalic regions.     

非整倍体胚胎发育中MSL复合体组分的RNA表达分析

MSL (male specific lethal) 复合体作为调节果蝇剂量补偿的关键元件，在非整倍体基因表达调控过程中发挥了重要作用．为深入探究非整倍体胚胎发育过程的分子调控机制，运用TSA-FISH (tyramide signal amplification- fluorescence in situ hybridization) 技术比较分析正常二倍体胚胎与常染色体三体胚胎中的MSL复合体组分基因表达模式，探究复合体重要组分在果蝇非整倍体胚胎不同时期的表达水平，以及亚细胞定位的变化．研究结果表明，在MSL复合体尚未组装的非整倍体胚胎发育早期，即母体表达阶段，多数复合体组分基因就已存在转录本水平上的差异，而这种表达差异主要源自于母体的非单倍体配子．随着发育的进行，染色体片段的增加导致基因组内剂量平衡发生变化，由此产生的反式剂量效应将引发MSL复合体各组分表达水平的差异变化，而这种差异将持续作用于后续非整倍体胚胎发育的各个时期．      

Control of neuronal fate by the Drosophila segmentation gene even-skipped

The central nervous system (CNS) contains a remarkable diversity of cell types. The molecular basis for generating this neuronal diversity is poorly understood. Much is known, however, about the regulatory genes which control segmentation and segment identity during early Drosophila embryogenesis. Interestingly, most of the segmentation and homoeotic genes in Drosophila, as well as many of their vertebrate homologues, are expressed during the development of the nervous system (for example, ref. 3). Are these genes involved in specifying the identity of individual neurons during neurogenesis, just as they specify the identity of cells during segmentation? We previously described the CNS expression of the segmentation gene fushi tarazu (ftz) and showed that ftz CNS expression is involved in the determination of an identified neuron. Here we show that another segmentation gene, even-skipped (eve), is expressed in a different but overlapping subset of neurons. Temperature-sensitive inactivation of the eve protein during neurogenesis alters the fate of two of these neurons. Our results indicate that the nuclear protein products of the eve and ftz segmentation genes are components of the mechanism controlling cell fate during neuronal development.