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Current Biology : CB Aug 2003
Topics: Animals; Drosophila Proteins; Membrane Proteins; Serine Endopeptidases
PubMed: 12906806
DOI: 10.1016/s0960-9822(03)00519-0 -
Annals of the New York Academy of... Apr 2005Here, we report the identification of proteins associated with the immune response of Drosophila by analyzing the hemolymph profiles after infection. Two-dimensional... (Comparative Study)
Comparative Study
Here, we report the identification of proteins associated with the immune response of Drosophila by analyzing the hemolymph profiles after infection. Two-dimensional difference gel electrophoresis was used to study the secretome in the hemolymph of Drosophila larvae. Shortly after induction with lipopolysaccharides, we identified 10 proteins, which we designated "Drosophila instantly released immune proteins". Infection with Micrococcus luteus or Saccharomyces cerevisiae induced 20 and 19 differential protein spots, respectively. Next to known immune proteins, new candidates that require further investigation were identified.
Topics: Animals; Drosophila Proteins; Drosophila melanogaster; Immunity, Innate; Proteomics
PubMed: 15891102
DOI: 10.1196/annals.1327.104 -
Archives of Biochemistry and Biophysics Mar 2021Cryptochromes, FAD-dependent blue light photoreceptors, undergo a series of electron transfer reactions after light excitation. Time-resolved optical spectroscopy was...
Cryptochromes, FAD-dependent blue light photoreceptors, undergo a series of electron transfer reactions after light excitation. Time-resolved optical spectroscopy was employed to investigate the pH dependence of all light-dependent reactions in the cryptochrome from fruit flies. Signal state formation experiments on a time scale of seconds were found to be strongly pH dependent, and formation of both anionic and neutral FAD radicals could be detected, with reaction rates increasing by a factor of ~2.5 from basic to neutral pH values. Additionally, the influence of the amino acid His378 was investigated in further detail: Two protein variants, DmCry H378A and H378Q, showed significantly reduced rate constants for signal state formation, which again differed at neutral and alkaline pH values. Hence, His378 was identified as an amino acid responsible for the pronounced pH dependence; however, this amino acid can be excluded as a proton donor for the protonation of the anionic FAD radical. Other conserved amino acids appear to alter the overall polarity of the binding pocket and thus to be responsible for the pronounced pH dependence. Furthermore, the influence of pH and other experimental parameters, such as temperature, glycerol or ferricyanide concentrations, on the intermediately formed FAD-tryptophan radical pair was explored, which deprotonates on a microsecond time scale with a clear pH dependence, and subsequently recombines within milliseconds. Surprisingly, the latter reaction showed no pH dependence; potential reasons are discussed. All results are reviewed in terms of the photoreceptor and potential magnetoreceptor functions of Drosophila cryptochrome.
Topics: Amino Acid Substitution; Animals; Cryptochromes; Drosophila Proteins; Drosophila melanogaster; Eye Proteins; Hydrogen-Ion Concentration; Mutation, Missense; Oxidation-Reduction; Protein Stability
PubMed: 33545100
DOI: 10.1016/j.abb.2021.108787 -
Cell Chemical Biology Jul 2016Cell-specific proteomics in multicellular systems and whole animals is a promising approach to understand the differentiated functions of cells and tissues. Here, we...
Cell-specific proteomics in multicellular systems and whole animals is a promising approach to understand the differentiated functions of cells and tissues. Here, we extend our stochastic orthogonal recoding of translation (SORT) approach for the co-translational tagging of proteomes with a cyclopropene-containing amino acid in response to diverse codons in genetically targeted cells, and create a tetrazine-biotin probe containing a cleavable linker that offers a way to enrich and identify tagged proteins. We demonstrate that SORT with enrichment, SORT-E, efficiently recovers and enriches SORT tagged proteins and enables specific identification of enriched proteins via mass spectrometry, including low-abundance proteins. We show that tagging at distinct codons enriches overlapping, but distinct sets of proteins, suggesting that tagging at more than one codon enhances proteome coverage. Using SORT-E, we accomplish cell-specific proteomics in the fly. These results suggest that SORT-E will enable the definition of cell-specific proteomes in animals during development, disease progression, and learning and memory.
Topics: Amino Acids; Animals; Azo Compounds; Biotin; Drosophila Proteins; Drosophila melanogaster; Female; Genetic Code; Mass Spectrometry; Molecular Probes; Molecular Structure; Protein Transport; Proteomics; Tetrazoles
PubMed: 27447048
DOI: 10.1016/j.chembiol.2016.05.018 -
Developmental Biology Jul 2022Cortical domains are characterized by spatially restricted polarity proteins. The pattern of cortical domains is dynamic and changes during cell differentiation and...
Cortical domains are characterized by spatially restricted polarity proteins. The pattern of cortical domains is dynamic and changes during cell differentiation and development. Although there is a good understanding for how the cortical pattern is maintained, e. g. by mutual antagonism, less is known about how the initial pattern is established, and its dynamics coordinated with developmental progression. Here we investigate the initial restriction of subapical marker proteins during the syncytial-cellular transition in Drosophila embryos. The subapical markers Canoe/Afadin, the complex ELMO-Sponge, Baz and Arm become initially restricted between apical and lateral domains during cellularization. We define the role of zygotic genome activation as a timer for subapical domain formation. Subapical markers remained widely spread in embryos treated with α-amanitin and became precociously restricted in mutant embryos with premature zygotic transcription. In contrast, remodeling of the nuclear division cycle without cytokinesis to a full cell cycle is not a prerequisite for subapical domain formation, since we observed timely subapical restriction in embryos undergoing an extra nuclear cycle. We provide evidence that earliest subapical markers ELMO-Sponge and Canoe are required for subapical accumulation of Baz. Supporting an important role of cortical F-actin in subapical restriction, we found that the formin Dia was required for Baz restriction, and its distribution depended on the onset of zygotic gene expression. In summary, we define zygotic transcription as a timer, to which subapical markers respond in a dia-dependent mechanism.
Topics: Animals; Drosophila; Drosophila Proteins; Formins; Morphogenesis; Zygote
PubMed: 35525304
DOI: 10.1016/j.ydbio.2022.04.012 -
PloS One Mar 2008The development of the Drosophila eye imaginal disc requires complex epithelial rearrangements. Cells of the morphogenetic furrow are apically constricted and this leads...
BACKGROUND
The development of the Drosophila eye imaginal disc requires complex epithelial rearrangements. Cells of the morphogenetic furrow are apically constricted and this leads to a physical indentation in the epithelium. Posterior to the furrow, cells start to rearrange into distinct clusters and eventually form a precisely patterned array of ommatidia. These morphogenetic processes include regulated changes of adhesion between cells.
METHODOLOGY/PRINCIPAL FINDINGS
Here, we show that two transmembrane adhesion proteins, Capricious and Tartan, have dynamic and complementary expression patterns in the eye imaginal disc. We also describe novel null mutations in capricious and double null mutations in capricious and tartan. We report that they have redundant functions in regulating the architecture of the morphogenetic furrow and ommatidial spacing.
CONCLUSIONS/SIGNIFICANCE
We conclude that Capricious and Tartan contribute to the adhesive properties of the cells in the morphogenetic furrow and that this regulated adhesion participates in the control of spacing ommatidial clusters.
Topics: Alleles; Animals; Drosophila; Drosophila Proteins; Membrane Proteins; Mutation; Pigment Epithelium of Eye
PubMed: 18350163
DOI: 10.1371/journal.pone.0001827 -
Biochemical and Biophysical Research... Dec 2003With the completion of the genome sequence of Drosophila melanogaster the importance of constructing a proteome map is to be considered. Therefore, with the application...
With the completion of the genome sequence of Drosophila melanogaster the importance of constructing a proteome map is to be considered. Therefore, with the application of recent advances in proteomic analysis approaches, a protein map of D. melanogaster larvae hemolymph proteins was obtained using 2-DE in the range of pH 3-10. After Coomassie colloidal detection of 289 spots, a total of 105 were excised from the gel and digested with trypsin. Identification was done based on a combination of MALDI-TOF/TOF MS and MS/MS spectra. The 99 proteins identified using this approach include a large number of metabolic enzymes, translational apparatus components, and structural proteins. Among these we emphasize the identification of proteins with molecular chaperone properties (heat shock proteins and PPIases) and protein spots involved in defense responses such as antioxidant and immunological defense mechanisms (thioredoxin, prophenoloxidase, and serine proteases), as well as in signal transduction pathways.
Topics: Amino Acid Sequence; Animals; Databases, Protein; Drosophila Proteins; Drosophila melanogaster; Electrophoresis, Gel, Two-Dimensional; Hemolymph; Larva; Molecular Sequence Data; Proteome; Proteomics; Sequence Analysis, Protein; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 14680800
DOI: 10.1016/j.bbrc.2003.10.156 -
International Journal of Molecular... Nov 2018Transcription factors (TFs) play essential roles in the transcriptional regulation of functional genes, and are involved in diverse physiological processes in living... (Review)
Review
Transcription factors (TFs) play essential roles in the transcriptional regulation of functional genes, and are involved in diverse physiological processes in living organisms. The fruit fly , a simple and easily manipulated organismal model, has been extensively applied to study the biological functions of TFs and their related transcriptional regulation mechanisms. It is noteworthy that with the development of genetic tools such as CRISPR/Cas9 and the next-generation genome sequencing techniques in recent years, identification and dissection the complex genetic regulatory networks of TFs have also made great progress in other insects beyond . However, unfortunately, there is no comprehensive review that systematically summarizes the structures and biological functions of TFs in both model and non-model insects. Here, we spend extensive effort in collecting vast related studies, and attempt to provide an impartial overview of the progress of the structure and biological functions of current documented TFs in insects, as well as the classical and emerging research methods for studying their regulatory functions. Consequently, considering the importance of versatile TFs in orchestrating diverse insect physiological processes, this review will assist a growing number of entomologists to interrogate this understudied field, and to propel the progress of their contributions to pest control and even human health.
Topics: Animals; Drosophila; Drosophila Proteins; Evolution, Molecular; Transcription Factors; Transcriptional Activation
PubMed: 30469390
DOI: 10.3390/ijms19113691 -
Journal of Proteomics Oct 2010Advances in mass spectrometry technology, high-throughput proteomics and genome annotations have resulted in significant increases in our molecular understanding of...
Advances in mass spectrometry technology, high-throughput proteomics and genome annotations have resulted in significant increases in our molecular understanding of sperm composition. Using improved separation and detection methods and an updated genome annotation, a re-analysis of the Drosophila melanogaster sperm proteome (DmSP) has resulted in the identification of 956 sperm proteins. Comparative analysis with our previous proteomic dataset revealed 766 new proteins and an updated sperm proteome containing a total of 1108 proteins, termed the DmSP-II. This expanded dataset includes additional proteins with predicted sperm functions and confirms previous findings concerning the genomic organization of sperm loci. Bioinformatic and protein network analyses demonstrated high quality and reproducibility of proteome coverage across three replicate mass spectrometry runs. The use of whole-cell proteomics in conjunction with characterized phenotypes, functional annotations and pathway information has advanced our systems level understanding of sperm proteome functional networks.
Topics: Animals; Chromatography, High Pressure Liquid; Databases, Protein; Drosophila Proteins; Drosophila melanogaster; Genome, Insect; Male; Mass Spectrometry; Phenotype; Proteome; Proteomics; Spermatozoa
PubMed: 20833280
DOI: 10.1016/j.jprot.2010.09.002 -
Doklady. Biochemistry and Biophysics May 2019We tested the activity of the PRE element from the Drosophila virilis genome, which is the homologue of the known Drosophila melanogasterbxdPRE element from the...
We tested the activity of the PRE element from the Drosophila virilis genome, which is the homologue of the known Drosophila melanogasterbxdPRE element from the regulatory region of the Ubx gene. It is easy to select unique primers to this element that do not occur in the Drosophila melanogaster genome. We showed that the studied PRE element causes a strong repression of the white marker gene upon insertion into the Drosophila melanogaster genome and interacts with the PcG proteins of the PRC1 and PhoRC complexes.
Topics: Animals; DNA Transposable Elements; Drosophila; Drosophila Proteins; Genome, Insect; Silencer Elements, Transcriptional
PubMed: 31012008
DOI: 10.1134/S1607672919010095