RESUMO
Orthodenticle (Otd)-related transcription factors are essential for anterior patterning and brain morphogenesis from Cnidaria to Mammals, and genetically underlie several human retinal pathologies. Despite their key developmental functions, relatively little is known regarding the molecular basis of how these factors regulate downstream effectors in a cell- or tissue-specific manner. Many invertebrate and vertebrate species encode two to three Otd proteins, whereas Drosophila encodes a single Otd protein. In the fly retina, Otd controls rhabdomere morphogenesis of all photoreceptors and regulates distinct Rhodopsin-encoding genes in a photoreceptor subtype-specific manner. Here, we performed a structure-function analysis of Otd during Drosophila eye development using in vivo rescue experiments and in vitro transcriptional regulatory assays. Our findings indicate that Otd requires at least three distinct transcriptional regulatory domains to control photoreceptor-specific rhodopsin gene expression and photoreceptor morphogenesis. Our results also uncover a previously unknown role for Otd in preventing co-expression of sensory receptors in blue vs. green-sensitive R8 photoreceptors. Sequence analysis indicates that many of the transcriptional regulatory domains identified here are conserved in multiple Diptera Otd-related proteins. Thus, these studies provide a basis for identifying shared molecular pathways involved in a wide range of developmental processes.
Assuntos
Diferenciação Celular/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Proteínas de Homeodomínio/genética , Células Fotorreceptoras de Invertebrados/citologia , Sequências Reguladoras de Ácido Nucleico/genética , Transcrição Gênica , Sequência de Aminoácidos , Animais , Sequência Conservada/genética , Proteínas de Drosophila/química , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/química , Dados de Sequência Molecular , Morfogênese/genética , Células Fotorreceptoras de Invertebrados/metabolismo , Regiões Promotoras Genéticas/genética , Rodopsina/genética , Rodopsina/metabolismo , Alinhamento de Sequência , Deleção de Sequência/genética , Homologia de Sequência de AminoácidosRESUMO
Low-temperature isomeric energies, structures, and properties of benzene-cyclohexane clusters are investigated via Monte Carlo simulations. The Monte Carlo strategy is first documented and then applied to (C(6)H(6))(C(6)H(12)) and (C(6)H(6))(C(6)H(12))(2) using four different potential energy surfaces. Results identify a single parallel-displaced dimer isomer. MP2 optimizations and frequency calculations support the Monte Carlo dimer structure and identify the van der Waals mode observed in vibronic spectra. Caloric simulations identify two temperatures where structural transitions occur and imply an experimental temperature below 10 K for dimers in cold supersonic expansions. The (C(6)H(6))(C(6)H(12))(2) studies identify eight independent trimer isomers: three form parallel-stacked (sandwich) arrangements with the two cyclohexane moieties related through a D(6)(h) transformation. The remaining five trimer isomers are trigonal, with no overall symmetry. Caloric studies indicate that the sandwich and trigonal isomeric classes coexist independently below 60 K, consistent with trimer vibronic spectra that contain two independent van der Waals progressions.