RESUMEN
In Drosophila, the dosage compensation complex (DCC) mediates upregulation of transcription from the single male X chromosome. Despite coating the polytene male X, the DCC pattern looks discontinuous and probably reflects DCC dynamic associations with genes active at a given moment of development in a salivary gland. To test this hypothesis, we compared binding patterns of the DCC and of the elongating form of RNA polymerase II (PolIIo). We found that, unlike PolIIo, the DCC demonstrates a stable banded pattern throughout larval development and escapes binding to a subset of transcriptionally active areas, including developmental puffs. Moreover, these proteins are not completely colocalized at the electron microscopy level. These data combined imply that simple recognition of PolII machinery or of general features of active chromatin is either insufficient or not involved in DCC recruitment to its targets. We propose that DCC-mediated site-specific upregulation of transcription is not the fate of all active X-linked genes in males. Additionally, we found that DCC subunit MLE associates dynamically with developmental and heat-shock-induced puffs and, surprisingly, with those developing within DCC-devoid regions of the male X, thus resembling the PolIIo pattern. These data imply that, independently of other MSL proteins, the RNA-helicase MLE might participate in general transcriptional regulation or RNA processing.
Asunto(s)
Cromosomas/metabolismo , Compensación de Dosificación (Genética)/fisiología , Drosophila/genética , Transcripción Genética , Animales , Animales Modificados Genéticamente , Proteínas de Drosophila/genética , Técnica del Anticuerpo Fluorescente , Regulación del Desarrollo de la Expresión Génica/fisiología , Larva/genética , Masculino , ARN Polimerasa II/genética , Factores de Transcripción/genética , Transgenes , Cromosoma X/metabolismoRESUMEN
Seven new alleles of the Broad-Complex gene of Drosophila melanogaster, which encodes a family of four zinc finger protein isoforms BR-C Z1, Z2, Z3 and Z4, were generated by transposase-induced mobilization of a P[Zw] element inserted in either the first intron downstream from the P165 promoter or the exon encoding the Z2-specific zinc finger domain. They were characterized by genetic complementation tests, molecular mapping and cytogenetic analysis of their effect on ecdysone-induced puffing and BR-C proteins binding to polytene chromosomes. Four mutations that correspond to three overlapping deletions and one tandem insertion of the P[Zw] element are located in the intron. They provide evidence that regulatory elements essential for a correct expression of the BR-C Z2 and BR-C Z3 transcripts are located within the intron downstream from the P165 promoter. Three mutations correspond to internal deletions of the locus and exhibit a complete loss of all BR-C(+) genetic functions in the complementation and cytogenetic tests. They thus provide well characterized new amorphic reference alleles of the BR-C gene. The precise cytogenetic location of more than 300 binding sites of BR-C proteins on larval salivary gland polytene chromosomes was determined by immunostaining using specific antibodies. Sites were found in big ecdysone inducible puffs, constitutively active small puffs as well as interbands. A complete list of the major sites on all four salivary gland polytene chromosomes of BR-C(+) larvae is presented.
Asunto(s)
Proteínas de Drosophila , Ecdisona/biosíntesis , Factores de Transcripción/genética , Dedos de Zinc , Animales , Cromosomas , Drosophila melanogaster/genética , Larva , Mutagénesis , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Factores de Transcripción/metabolismoRESUMEN
The properties of heterochromatin from polytene chromosomes of the malaria mosquito Anopheles messeae Fal. and A. atroparvus V. Tiel. were studied by various methods of differential staining and by hybridization in situ with two repetitive DNA sequences of Drosophila melanogaster. In malaria mosquito, the heterochromatin was heterogeneous. Two forms of alpha-heterochromatin were revealed: pericentromeric and intercalary heterochromatin, which is localized within the internal chromosome regions.