The Drosophila homolog of the mammalian imprint regulator, CTCF, maintains the maternal genomic imprint in Drosophila melanogaster

<p>Abstract</p> <p>Background</p> <p>CTCF is a versatile zinc finger DNA-binding protein that functions as a highly conserved epigenetic transcriptional regulator. CTCF is known to act as a chromosomal insulator, bind promoter regions, and facilitate long-range chromatin interactions. In mammals, CTCF is active in the regulatory regions of some genes that exhibit genomic imprinting, acting as insulator on only one parental allele to facilitate parent-specific expression. In <it>Drosophila</it>, CTCF acts as a chromatin insulator and is thought to be actively involved in the global organization of the genome.</p> <p>Results</p> <p>To determine whether CTCF regulates imprinting in <it>Drosophila</it>, we generated <it>CTCF </it>mutant alleles and assayed gene expression from the imprinted <it>Dp(1;f)LJ9 </it>mini-X chromosome in the presence of reduced <it>CTCF </it>expression. We observed disruption of the maternal imprint when <it>CTCF </it>levels were reduced, but no effect was observed on the paternal imprint. The effect was restricted to maintenance of the imprint and was specific for the <it>Dp(1;f)LJ9 </it>mini-X chromosome.</p> <p>Conclusions</p> <p>CTCF in <it>Drosophila </it>functions in maintaining parent-specific expression from an imprinted domain as it does in mammals. We propose that <it>Drosophila </it>CTCF maintains an insulator boundary on the maternal X chromosome, shielding genes from the imprint-induced silencing that occurs on the paternally inherited X chromosome.</p> <p>See commentary: <url>http://www.biomedcentral.com/1741-7007/8/104</url></p

“Where, O Death, Is Thy Sting?” A Brief Review of Apoptosis Biology

Apoptosis was a term introduced in 1972 to distinguish a mode of cell death with characteristic morphology and apparently regulated, endogenously driven mechanisms. The effector processes responsible for apoptosis are now mostly well known, involving activation of caspases and Bcl2 family members in response to a wide variety of physiological and injury-induced signals. The factors that lead of the decision to activate apoptosis as opposed to adaptive responses to such signals (e.g. autophagy, cycle arrest, protein synthesis shutoff) are less well understood, but the intranuclear Promyelocytic Leukaemia Body (PML body) may create a local microenvironment in which the audit of DNA damage may occur, informed by the extent of the damage, the adequacy of its repair and other aspects of cell status

Effects of a defective ERAD pathway on growth and heterologous protein production in Aspergillus niger

Endoplasmic reticulum associated degradation (ERAD) is a conserved mechanism to remove misfolded proteins from the ER by targeting them to the proteasome for degradation. To assess the role of ERAD in filamentous fungi, we have examined the consequences of disrupting putative ERAD components in the filamentous fungus Aspergillus niger. Deletion of derA, doaA, hrdC, mifA, or mnsA in A. niger yields viable strains, and with the exception of doaA, no significant growth phenotype is observed when compared to the parental strain. The gene deletion mutants were also made in A. niger strains containing single- or multicopies of a glucoamylase–glucuronidase (GlaGus) gene fusion. The induction of the unfolded protein response (UPR) target genes (bipA and pdiA) was dependent on the copy number of the heterologous gene and the ERAD gene deleted. The highest induction of UPR target genes was observed in ERAD mutants containing multiple copies of the GlaGus gene. Western blot analysis revealed that deletion of the derA gene in the multicopy GlaGus overexpressing strain resulted in a 6-fold increase in the intracellular amount of GlaGus protein detected. Our results suggest that impairing some components of the ERAD pathway in combination with high expression levels of the heterologous protein results in higher intracellular protein levels, indicating a delay in protein degradation

Eye Development under the control of SRp55/B52-Mediated Alternative Splicing of eyeless

The genetic programs specifying eye development are highly conserved during evolution and involve the vertebrate Pax-6 gene and its Drosophila melanogaster homolog eyeless (ey). Here we report that the SR protein B52/SRp55 controls a novel developmentally regulated splicing event of eyeless that is crucial for eye growth and specification in Drosophila. B52/SRp55 generates two isoforms of eyeless differing by an alternative exon encoding a 60-amino-acid insert at the beginning of the paired domain. The long isoform has impaired ability to trigger formation of ectopic eyes and to bind efficiently Eyeless target DNA sequences in vitro. When over-produced in the eye imaginal disc, this isoform induces a small eye phenotype, whereas the isoform lacking the alternative exon triggers eye over-growth and strong disorganization. Our results suggest that B52/SRp55 splicing activity is used during normal eye development to control eye organogenesis and size through regulation of eyeless alternative splicing

Catalase enzyme in mitochondria of Saccharomyces cerevisiae

Catalase and superoxide dismutase activities have been explored in the yeast Saccharomyces cerevisiae during batchwise growth experiment. During the diauxic growth in YPD medium high Ys values were obtained (0.415 - 0.423) and correlation between the total activities of both enzymes has been found. A mitochondrial fraction from three type strains of Saccharomyces cerevisiae has been isolated. The purity of this fraction was proved through different enzyme assays: hexokinase, glucose-6-phosphate dehydrogenase, D-amino acid oxidase, isocitric lyase, succinate dehydrogenase. Then the catalase, peroxidase, Mn and Cu/Zn superoxide dismutase activities were evaluated in the mitochondrial fraction. Polyacrylamide gel electrophoresis separations allowed to identify a mitochondrial catalase as a band of 0.239 Rm value. It differed from the two catalase specific bands with Rm values 0.218 and 0.257 obtained from the crude extract. It was proved that the three catalase proteins are charge isomers. A positive correlation between the activity of mitochondrial catalase and Mn superoxide dismutase also takes place. Molecular weight of mitochonrial catalase protein has been determined as 240 kD

Catalase enzyme in mitochondria of Saccharomyces cerevisiae

Catalase and superoxide dismutase activities have been explored in the yeast Saccharomyces cerevisiae during batchwise growth experiment. During the diauxic growth in YPD medium high Ys values were obtained (0.415 - 0.423) and correlation between the total activities of both enzymes has been found. A mitochondrial fraction from three type strains of Saccharomyces cerevisiae has been isolated. The purity of this fraction was proved through different enzyme assays: hexokinase, glucose-6-phosphate dehydrogenase, D-amino acid oxidase, isocitric lyase, succinate dehydrogenase. Then the catalase, peroxidase, Mn and Cu/Zn superoxide dismutase activities were evaluated in the mitochondrial fraction. Polyacrylamide gel electrophoresis separations allowed to identify a mitochondrial catalase as a band of 0.239 Rm value. It differed from the two catalase specific bands with Rm values 0.218 and 0.257 obtained from the crude extract. It was proved that the three catalase proteins are charge isomers. A positive correlation between the activity of mitochondrial catalase and Mn superoxide dismutase also takes place. Molecular weight of mitochonrial catalase protein has been determined as 240 kD