43 research outputs found

    A Cytosine Methyltransferase Homologue Is Essential for Sexual Development in Aspergillus nidulans

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    Background: The genome defense processes RIP (repeat-induced point mutation) in the filamentous fungus Neurospora crassa, and MIP (methylation induced premeiotically) in the fungus Ascobolus immersus depend on proteins with DNA methyltransferase (DMT) domains. Nevertheless, these proteins, RID and Masc1, respectively, have not been demonstrated to have DMT activity. We discovered a close homologue in Aspergillus nidulans, a fungus thought to have no methylation and no genome defense system comparable to RIP or MIP. Principal Findings: We report the cloning and characterization of the DNA methyltransferase homologue A (dmtA) gene from Aspergillus nidulans. We found that the dmtA locus encodes both a sense (dmtA) and an anti-sense transcript (tmdA). Both transcripts are expressed in vegetative, conidial and sexual tissues. We determined that dmtA, but not tmdA, is required for early sexual development and formation of viable ascospores. We also tested if DNA methylation accumulated in any of the dmtA/tmdA mutants we constructed, and found that in both asexual and sexual tissues, these mutants, just like wild-type strains, appear devoid of DNA methylation. Conclusions/Significance: Our results demonstrate that a DMT homologue closely related to proteins implicated in RIP and MIP has an essential developmental function in a fungus that appears to lack both DNA methylation and RIP or MIP. It remains formally possible that DmtA is a bona fide DMT, responsible for trace, undetected DNA methylation that i

    Highlights of the DNA cutters:a short history of the restriction enzymes

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    In the early 1950ā€™s, ā€˜host-controlled variation in bacterial virusesā€™ was reported as a non-hereditary phenomenon: one cycle of viral growth on certain bacterial hosts affected the ability of progeny virus to grow on other hosts by either restricting or enlarging their host range. Unlike mutation, this change was reversible, and one cycle of growth in the previous host returned the virus to its original form. These simple observations heralded the discovery of the endonuclease and methyltransferase activities of what are now termed Type I, II, III and IV DNA restriction-modification systems. The Type II restriction enzymes (e.g. EcoRI) gave rise to recombinant DNA technology that has transformed molecular biology and medicine. This review traces the discovery of restriction enzymes and their continuing impact on molecular biology and medicine

    Restriction and modification in Bacillus subtilis: DNA methylation potential of the related bacteriophages Z, SPR, SP beta, phi 3T, and rho 11.

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    The DNA methylation capacity and some other properties of the related temperate Bacillus subtilis phages Z, SPR, SP beta, phi 3T, and rho 11 are compared. With phage mutants affected in their methylation potential, we show that phage-coded methyltransferase genes are interchangeable among the phages studied. DNA/DNA hybridization experiments indicate that phage methyltransferase genes are structurally related, whereas no such relationship is observed to a bacterial gene, specifying a methyltransferase with the same specificity

    Restriction and Modification in Bacillus subtilis: Gene Coding for a BsuR-Specific Modification Methyltransferase in the Temperate Bacteriophage Ī¦3T

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    The resistance of Ī¦3T DNA to degradation by the restriction enzyme BsuR or its isoschizomer HaeIII is due to obligatory modification of such DNA. Biochemical and genetical experiments indicate that Ī¦3T codes for a methyltransferase, which methylates Ī¦3T DNA itself or heterologous DNA at target sites 5ā€²-GG(*)CC
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